Drug treatment of overactive bladder

ABSTRACT

Provided are methods of treating an overactive bladder in a patient which include: administering a Myosin II ATPase inhibitor compound; or administering an X group and Y group substituted (3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2, 3b] quinolin-4-one) compound of Formula 1 or administering an X group and Y group substituted (3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b] quinolin-4-one) compound of Formula II; or administering pharmaceutically-acceptable salts, racemic mixtures, enantiomers, or prodrugs of said compounds, useful in their active form as inhibitors of Myosin H ATPase related to over-active bladder. Optionally the compounds are administered intervesicularly into the bladder. Also provided are pharmaceutical compositions comprising said compounds useful in their active form, as methods of treating a patient suffering from an over-active bladder related to inhibition of Myosin II ATPase. These pharmaceutical compositions also may contain one or more other compounds useful in their active form, as methods of treating a patient suffering from an over-active bladder.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit to priority of U.S. ProvisionalApplication with Ser. No. 61/487,329 filed May 18, 2011.

GOVERNMENT FUNDING

The U.S. Government has a paid-up license in this invention and theright in limited circumstances to require the patent owner to licenseothers on reasonable terms as provided for by terms of research grantcontract number 5RO1DK077116 awarded by the National Institutes ofHealth (NIH).

FIELD OF THE INVENTION

The present invention relates to methods of treatment of over activebladder using Myosin ATPase inhibitor compounds, including substituted3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-onecompounds which in their active form can inhibit Myosin II ATPase.

BACKGROUND OF THE INVENTION

Lower urinary tract (LUT) dysfunction can include an overactive bladder(OAB). The bladder's smooth muscle is referred to as the detrusormuscle. The main symptoms of an overactive bladder are an increasedday-time and night-time frequency of urination, an increased frequencyof the urge to urinate and a reduced ability to control urination. OABis increasing in the ageing population and there are fewhighly-effective or tolerable treatments (Kelleher C J, Kreder K J,Pleil A M, Burgess S M, Reese P R, “Health-related quality of life ofpatients receiving extended-release tolterodine for overactive bladder”Am J Manag. Care 2002; 8: S608-15).

A cause of OAB may be a frequent or an excessive release ofacetylcholine from cholinergic nerves innervating or terminating nearthe bladder smooth muscle. A common pharmaceutical therapy for OAB isadministration of an anti-muscarinic drug to block the excessiveacetylcholine from binding to acetylcholine receptors. Unfortunately,such anti-muscarinic therapy has only a 65-75% efficacy in treatingmajor symptoms of overactive bladder (OAB) in a patient, and its use islimited due to drug side effects such as blurred vision, dry mouth andconstipation. (Zhang, X, Kuppam, D S R, Melman, A, DiSanto, M E. “Invitro and In vivo relaxation of urinary bladder smooth muscle by theselective Myosin II inhibitor, blebbistatin” BJU Internat. e-publication2010, journal publication, 2011; Volume 107, Issue 2, pages 310-317).About 75% of the OAB patients discontinue taking an anti-muscarinic drugto treat OAB because the patient finds such side effects intolerable.

Various types of drugs for a treatment of OAB include using α-adrenergicantagonists, β-adrenoceptor agonists, membrane channel activitymodulators, phosphodiesterase inhibitors, and prostaglandin-synthesisinhibitors (Andersson K E, Chapple C R, Cardozo L et al.“Pharmacological treatment of overactive bladder: report from theInternational Consultation on Incontinence” Curr Opin Urol 2009; 19:380-94). Some pharmaceutical drug therapies for OAB modify the myogenicpathway of the bladder's smooth muscle (detrusor muscle) (Yoshimura N,Kaiho Y, Miyazato M et al. “Therapeutic receptor targets for lowerurinary tract dysfunction.” Naunyn Schmiedebergs Arch Pharmacol 2008;377: 437-48). Myogenic pathways are believed to be an important triggerfor detrusor muscle contraction and relaxation cycling. In smooth musclethere are changes in cell membrane tension during the muscle contractionand relaxation cycle. The stretch declines during muscle contraction andincreases during muscle relaxation. Increased membrane tension opensmembrane ion channels whose conductance depolarizes the celltransmembrane potential. The change in transmembrane potential increasescytoplasmic calcium ion levels and other biochemical processes whichstimulate muscle contraction. Contraction relieves membrane tension andthe processes reverse during muscle relaxation when membrane tension hasagain increased due to muscle lengthening. Different smooth muscleorgans vary in degree, frequency and time profile of their contractions.The response of different smooth muscles to drugs is unpredictable. Itis appreciated that there is serial and parallel biochemical signalprocessing and often with positive and negative feedback controls. Also,there can be changes in genetic expression of smooth muscle duringdevelopment and aging. Endocrine and exocrine processes also modulatesmooth muscle function. It is not obvious how to develop an effectiveand tolerable pharmaceutical treatment for an over-active bladder smoothmuscle.

SUMMARY OF THE INVENTION

According to the invention there is disclosed a method of treating anoveractive bladder in a patient, which method comprises: administeringto said patient an effective amount of a Myosin II ATPase inhibitorcompound, or a pharmaceutically-acceptable salt, a racemic mixture, anenantiomer thereof; optionally administering a drug selected from thegroup consisting of a PDE5 inhibitor drug and an antimuscarininc drug,and optionally administering to the bladder directly or intervesicularyan effective amount the a Myosin II ATPase inhibitor compound, apharmaceutically-acceptable salt, a racemic mixture, an enantiomer, or aprodrug thereof, to the overactive bladder of the patient.

Further according to the invention, the invention also providespharmaceutical compositions comprising an effective amount of a MyosinII ATPase inhibitor compound, useful in its active form, as a method oftreating an overactive bladder in a patient.

Still further according to the invention, there is also disclosed amethod of treating an overactive bladder in a patient, which methodcomprises administering to said patient an effective amount of a MyosinII ATPase inhibitor compound of Formula (1) or apharmaceutically-acceptable salt, a racemic mixture, an enantiomer, or aprodrug

thereof,

wherein: X is selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent W,the substituent W selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—C₁₋₆)alkyl, acetamido, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, and optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent Q, the substituent Q selectedfrom the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic,(C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; wherein Y isselected from the group consisting of hydrogen, methyl, hydroxy, fluoro,chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a carbon atom with a substituent Z, thesubstituent Z selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andoptionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent T, the substituent T selected from thegroup consisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, (C₄₋₁₀)aryl, napthyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl. Optionally said method,comprises: administering a compound of formula I having an IC₅₀ of about0.0001 to about 50 micromolar for inhibiting a rat bladder muscle stripcontraction in vitro; optionally administering a drug selected from thegroup consisting of a PDE5 inhibitor drug and an antimuscarininc drug;and optionally administering to the bladder directly or intervesicularyan effective amount of a Myosin II ATPase inhibitor compound of formulaI, a pharmaceutically-acceptable salt, a racemic mixture, an enantiomer,or a prodrug thereof. Yet further according to the invention, a methodof treating an overactive bladder in a patient includes administering tosaid patient an effective amount of a Myosin II ATPase inhibitorcompound of formula (II), or a pharmaceutically-acceptable salt, aracemic mixture, an enantiomer, or a prodrug thereof,

wherein: X is selected from the group consisting of hydrogen, methyl,hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino, methylamino,aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent W,the substituent W selected from the group consisting of methyl,hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino, methylamino,aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,acetamido, cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl,(C₄₋₁₀)aryl, napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,(C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl, valinyl, leucinyl,isolecinyl, prolinyl, methionyl, phenyklalanyl, tyrosinyl, trytophanyl,threoninyl, cystinyl, asparginyl, glutamyl, lysinyl, histidinyl,arginyl, aspartyl, and glutamyl, and optionally substituted at a primarynitrogen atom or at a secondary nitrogen atom with a substituent Q, thesubstituent Q selected from the group consisting of methyl, (C₁₋₆)alkyl,cyclopropyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl,(C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, benzyl, napthyl,(C₄₋₈)heterocyclic, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl;wherein Y is selected from the group consisting of hydrogen, methyl,hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino, methylamino,aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,(C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent Z,the substituent Z selected from the group consisting of methyl,hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino, methylamino,aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,(C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, and optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent T, the substituent T selectedfrom the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, benzyl, (C₄₋₁₀)aryl, napthyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; and whereinan atom of X and an atom of Y are in a chemical bond that forms a ringfrom X and Y; wherein said ring is ortho-fused to substituent positions6 and 7 of a ring of Formula (II). Optionally said method, comprises:administering a compound of formula II having an IC₅₀ of about 0.0001 toabout 50 micromolar for inhibiting a rat bladder muscle stripcontraction in vitro; optionally administering a drug selected from thegroup consisting of a PDE5 inhibitor drug and an antimuscarininc drug;and optionally administering to the bladder directly or intervesicularyan effective amount of a Myosin II ATPase inhibitor compound of formulaII, a pharmaceutically-acceptable salt, a racemic mixture, anenantiomer, or a prodrug thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a tracing of the change in tension (contraction) versustime of a rat bladder smooth muscle strip mounted in a tissue bathpre-contracted with 60 millimolar KCL (potassium chloride) and thenrelaxed using 1 uM, then 5 uM and then 10 uM BLEB. The abbreviation uMmeans micromolar. The abbreviation BLEB means racemic blebbistatin. Aracemic mixture of blebbistatin [(±) of BLEB] was used in all studiesunless otherwise noted. The x-axis represents time in minutes (min)while the y-axis represents force (mg). Maximal response topre-contraction stimulus was taken as 100%, while the relaxant effect ofcumulative concentrations (1-10 μM) of BLEB was evaluated as apercentage of this response.

FIG. 2 shows a tracing of the change in tension (contraction) versustime of a rat bladder smooth muscle strip mounted in a tissue bathpre-contracted with 1 uM carbacol (CC) and then relaxed using 1 uM, then5 uM and then 10 uM BLEB (racemic blebbistatin). The x-axis representstime in minutes (min) while the y-axis represents force (mg). Maximalresponse to pre-contraction stimulus was taken as 100%, while therelaxant effect of cumulative concentrations (1-10 μM) of BLEB wasevaluated as a percentage of this response.

FIG. 3 shows a tracing of the change in tension (contraction) versustime of a spontaneously slowly contracting rat bladder smooth musclestrip mounted in a tissue bath pre-contracted with 1 uM carbacol (CC)which increased its baseline tension and the magnitude of its phasiccontractions. The rat bladder smooth muscle strip was then exposed to 1uM (+)BLE, then 5 uM (+BLE), 10 uM (+)BLE. Here the abbreviation (+)BLEand the abbreviation (+) BLEB shall both mean the (+)-blebbistatin or(R)-blebbistatin enantiomer. The rat bladder smooth muscle strip wasthen exposed to 10 uM (±) BLE. Here the abbreviation (±) BLE and theabbreviation (±) BLEB shall both mean racemic blebbistatin which can bewritten as (±)-blebbistatin. Racemic blebbistatin caused a significantdecrease in both the baseline and peak phasic force of contraction. Theenantiomer (+)-blebbistatin is relatively low potency (has a very highIC50) as an inhibitor of Myosin ATPase and was used as Control in someexperiments of the present invention. The x-axis represents time inminutes (min) while the y-axis represents force (mg). Maximal responseto pre-contraction stimulus was taken as 100%, while the relaxant effectof cumulative concentrations of 1-10 uM (+) BLE and 10 uM (±) BLE wasevaluated as a percentage of this response.

FIG. 4A shows a tracing of the change in tension (contraction) versustime of a rat bladder smooth muscle strip mounted in a tissue bathpre-contracted with 1 uM carbacol (CC). After several washes withoutdrug to return tension to the basal state, then the detrusor muscle(bladder smooth muscle) was pre-incubated with 10 μM BLEB (racemicblebbistatin) for 30 minutes (min). The tissue was pre-contracted againwith 1 uM carbacol while in the presence of 10 uM BLEB.

FIG. 4B shows a summary graph for the data of FIG. 4A. **=p<0.01 vs withBLEB (racemic blebbistatin). (n=4 strips obtained from 4 differentanimals).

FIG. 5A shows the tension (contraction) development and relaxationversus time of a rat bladder smooth muscle strip mounted in a tissuebath and pre-contracted with 60 mM KCl, during episodes of increasingelectric field stimulation (EFS) frequency (2 Hz to 4 Hz, to 8 Hz and to16 Hz). Hz is an abbreviation for Hertz which means frequency incycles/second. After several washes without drug to return tension tothe basal state, then the detrusor muscle (bladder smooth muscle) waspre-incubated with 10 μM BLEB (racemic blebbistatin) for 30 minutes(min) and then the EFS protocol was repeated.

FIG. 5B is a summary graph for the contraction data of FIG. 5A at EFSfrequencies 2 Hz, 4 Hz, 8 Hz and 16 Hz with or without 10 uM BLEB(racemic blebbistatin) as a percentage of maximal contraction induced by60 mM KCl. (*=p<0.05 vs with BLEB); (**=p<0.01 vs with BLEB). (n=4strips obtained from 4 different animals).

FIG. 6A shows a representative force tracing versus time of BLEB's(racemic blebbistatin's) in vitro relaxant effect on endothelin-1pre-contracted Human bladder. Human bladder (base bladder near bladderneck) strips were mounted in a tissue bath, pre-contracted with 20 nMendothelin-1 (ET-1) and then treated with cumulative doses of BLEB. Thex-axis represents time in minutes (min) while the y-axis representsforce (mg).

FIG. 6B is a summary graph for the data of FIG. 6A. The maximal responseto endothelin-1 (ET-1) was taken as 100%, while the relaxant effect ofBLEB (racemic blebbistatin) was evaluated as a percentage of thisresponse. Values are expressed as mean±SEM. (n=4 different strips).

FIG. 7A shows an extended time tracing of spontaneous contractionactivity versus time of rat bladder strips mounted in a tissue bath andthen 10 μM BLEB (racemic blebbistatin) was added and observed for 60minutes. The x-axis represents time in minutes (min) while the y-axisrepresents force (mg). The x-axis represents time (seconds) while they-axis represents force (mg).

FIG. 7B shows zoomed in tracings of spontaneous contraction activityversus time of rat bladder strips mounted in a tissue bath and then 10μM BLEB (racemic blebbistatin) was added and observed for 60 minutes.The x-axis represents time in minutes (min) while the y-axis representsforce (mg). The x-axis represents time (seconds) while the y-axisrepresents force (mg). before BLEB treatment while lower panel is aftertreatment.

FIG. 8A shows urodynamics of a bladder in an awake rat. After 90 minutesof saline infusion into bladder, there is an infusion of 250 nanomolarBLEB (racemic blebbistatin). Upper panels depict intravesical pressure(x-axis=minutes time and y-axis=pressure in cm H₂O. Lower panels givevolume and frequency of micturation (urine voiding of bladder). Thex-axis represents time in minutes (min) while the y-axis representsvolume (ml). Left side panels are control experiments with salinevehicle infusion while right side panels are after infusion of BLEB.

FIG. 8B shows urodynamics of an overactive bladder in an awake rat.The * represents when urine collection tube had to be emptied as it wasfull. Upper panel is a control experiment with saline while lower panelis urodynamics of the bladder after an infusion of 250 nanomolar BLEB(racemic blebbistatin). Volume and frequency of micturation by the ratwith the overactive bladder decrease due to BLEB. The x-axis representstime (min) while the y-axis represents volume (ml).

DETAILED DESCRIPTION OF THE INVENTION

A patient with an overactive bladder (a patient with OAB or an OABpatient) has an increased urge to urinate and less control as to whenurination shall occur compared to a normal person. It is an importantobject of the present invention to provide a therapeutically effectivemethod of treating an overactive bladder in a patient, so that the OABpatient can have a normal degree of voluntarily control their bladder.To be therapeutically effective, the present invention method oftreating an overactive bladder in a patient comprises administering tothe patient an effective amount of a Myosin II ATPase inhibitorcompound.

During experiments for the present invention, it was surprisingly foundthat the Myosin II ATPase inhibitor blebbistatin reduced tonic andphasic contractions in adult rat bladder smooth muscle strips induced bypotassium chloride, carbacol (carbamylcholine), and electric fieldstimulation. In follow-up OAB animal model experiments for the presentinvention, a blebbistatin composition was instilled (administered) by acatheter into the over-active bladder of an adult living rat.

Myosin II ATPase is an enzyme associated with the contraction process ofsmooth muscle. By genetic and chromatographic methods, there arepresently four Myosin II ATPase isoforms that have been detected insmooth muscle. The differences in their polypeptide sequences anddifferences in their conformational structures have unknownconsequences. Isoforms of a protein can be produced by related genes, ormay arise from the same gene by alternative splicing. Some isoforms arecaused by single-nucleotide polymorphisms or SNPs, small geneticdifferences between alleles of the same gene. SNPs can occur at specificindividual nucleotide positions within a gene. In smooth muscle thereare at least four slice variants from a single Myosin ATPase gene. Inaddition to the heavy chains there are 2 light chains and at least foursplice variants of the light chains (Aguilar, H. N.: Xiao, S: Knoll, A.H.; Yuan, X (2010) “Physiological pathways and molecular mechanismsregulating uterine contractility” Human Reproduction Update: 16 (6)725).

Blebbistatin is a small organic molecule with chemical structure name:3a-hydroxy-6-methyl-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-one,that was discovered by vitro testing for an inhibitor of a non-muscleMyosin II ATPase using IC₅₀ measurements as a criteria (Straight A F,Cheung A, Limouze J et al. “Dissecting temporal and spatial control ofcytokinesis with a Myosin II Inhibitor” Science 2003; 299: 1743-7). AnIC₅₀ measurement is a determination of the concentration of an inhibitorthat causes a 50% inhibition of a biological process such as an enzymeactivity, and the IC₅₀ can have units of concentration, such as uM(micromolar). The potency of an inhibitor is inversely-related to itsIC₅₀ value. The Myosin II ATPase inhibitor, blebbistatin, has beenreported to have significantly different IC₅₀ values in vitro indifferent tissues, such as for example, in rabbit striated skeletalmuscle bebbistatin inhibits the Myosin II ATPase with an IC₅₀ value of0.5 uM; in pig cardiac muscle bebbistatin inhibits the Myosin II ATPasewith an IC₅₀ value of 1.2 uM); and in turkey smooth muscle bebbistatininhibits the Myosin II ATPase with an IC₅₀ value of 79.6 uM). (LimouzeJ, Straight A F, Mitchison T, Sellers J R. “Specificity of blebbistatin,an inhibitor of Myosin II.” J Muscle Res Cell Motil 2004; 25:337-41).Furthermore, Ekman et al. reported that 10 uM blebbistatin did not blockan adult mouse bladder smooth muscle contraction when the tissue isdepolarized using potassium chloride. However, 10 uM blebbistatin didinhibit the bladder contraction of a newborn mouse. The new-born mousebladder was found to predominantly express a different Myosin II ATPaseisoform known as the non-muscle Myosin II ATPase isoform (Ekman M,Fagher K, Wede M, Stakeberg K, Amer A. “Decreased phosphatase activity,increased Ca2+ sensitivity, and myosin light chain phosphorylation inurinary bladder smooth muscle of newborn mice” J Gen Physiol 2005; 125:187-96).

It was surprisingly found that a pharmaceutical composition of theMyosin ATPase inhibitor blebbistatin, based upon measured urodynamicparameters, reduced the over-activity of the adult rat bladder in vivo.During other contraction experiments on isolated human bladder stripsfor the present invention it was surprisingly found that the Myosin IIATPase inhibitor blebbistatin completely blocked the strong contractioninduced by endothelin-1 in human bladder tissue strips.

When an effective amount of a Myosin II ATPase inhibitor compound inaccordance with the present invention has been administered to an OABpatient, the patient experiences one or more of the followingtherapeutic effects: a reduced urge to urinate; an increased degree ofvoluntary control of their bladder; a less frequent urination; and alarger urination volume per urination as compared to prior to atreatment with an effective amount of a Myosin ATPase inhibitorcompound.

In treating an OAB patient, the doctor of an OAB patient can applyexperience learned from helping other OAB patients and can refer toclinical and preclinical test reports on the treatment and observationof an OAB patient. Administered to an OAB patient, an effective amountof a Myosin II ATPase inhibitor compound will be observed to reduce thefrequency of urination by an OAB patient. One can give an OAB patientbeginning treatment for the first time, a small amount of a Myosin IIATPase inhibitor compound and evaluate its clinical effectiveness.Timing and measuring volume of urine collection can be used to determinewhen the OAB patient is being administered an effective amount of theMyosin II ATPase inhibitor. In addition the OAB patient can inform thedoctor about how well the medicine is working, as for example,preferably the frequency of urination by an OAB patient would be reducedso that the patient would not have to wake up more than once duringtheir night time sleep. Secondly, increased voluntary control ofurination would be another means for knowing if an effective amount ofthe Myosin II ATPase inhibitor had been administered. Again the OABpatient can indicate when they believe they are being administered aneffective amount of the Myosin II ATPase inhibitor.

The invention provides a method of treating an overactive bladder in apatient, the method comprising administering to the patient an effectiveamount of a Myosin II ATPase inhibitor compound, useful in its activeform, for example comprising a3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-oneas represented by structural Formula 1, or a pharmaceutically-acceptablesalt, racemic mixture, enantiomer, or prodrug thereof,

wherein X is selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent W,the substituent W selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, acetamido, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent Q, the substituent Q selectedfrom the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic,(C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; wherein Y isselected from the group consisting of hydrogen, methyl, hydroxy, fluoro,chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a carbon atom with a substituent Z, thesubstituent Z selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andoptionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent T, the substituent T selected from thegroup consisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, (C₄₋₁₀)aryl, napthyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl.

Provided is a test for a formula I or a formula II compound of thepresent invention. The test allows for a determination of the IC₅₀ ofthe compound for an inhibition of a rat bladder smooth musclecontraction in vitro. The IC₅₀ test is described in detail beforeEXAMPLE 1 of the written description and is entitled “IC₅₀ of a Compoundof the Invention for Inhibition of Contraction of a Rat Bladder Strip”.Also this Test may be referred to herein as a test measuring “inhibitionof contraction of rat bladder muscle strip in vitro” and the compound'sIC₅₀ data may be called “rat bladder strip contraction IC₅₀ data”.

When a Myosin II ATPase inhibitor compound of formula 1 and of formulaII is “in an active form”, this means the compound of the invention isactive and can inhibit Myosin II ATPase and can be tested in vitro on arat bladder to determine its IC₅₀ for inhibition of a contraction ofsaid bladder. A Myosin II ATPase inhibitor compound of formula 1 and offormula II “in an active form” can have an IC₅₀ value between about0.001 to about 50 micromolar based on inhibiting a contraction of a ratbladder muscle strip in vitro. A Myosin II ATPase inhibitor compound offormula 1 and of formula II in its active form can have an IC50 of about0.0001 to about 20 micromolar, the IC50 based on inhibiting a ratbladder muscle strip contraction in vitro.

The present invention includes methods of treatment of an overactivebladder of a patient which comprise administering a compound which is aMyosin II ATPase inhibitor, wherein the compound a prodrug. A prodrug ofthe present invention is a compound which may not be active as aninhibitor of Myosin II ATPase. A compound of the invention in itsprodrug form has a chemical group modification. When the chemical groupmodification is removed from the prodrug, then the compound of theinvention may be “in its active form” as an inhibitor of Myosin IIATPase. For example a prodrug of Formula I and II compounds can have anester group as a chemical group modification, the chemical groupmodification located as an optional substituent of X or of Y or as achemical modification group of the hydroxy group located at position 3aof a Formula I or a Formula II compound of the invention. When the estergroup no longer present on the compound, then compound may be “in itsactive form”.

Some Myosin II ATPase inhibitor compounds of formula 1 and formula II intheir active form have an IC₅₀ values between about 0.001 to about 50micromolar micromolar based on inhibiting a contraction of a rat bladdermuscle strip in vitro. Preferably, the Myosin II ATPase inhibitorcompounds of formula 1 and formula II in their active form have an IC₅₀values between about 0.001 to about 20 micromolar micromolar based oninhibiting a contraction of a rat bladder muscle strip in vitro; morepreferably Myosin II ATPase inhibitor compounds of formula 1 and formulaII in their active form, have an IC₅₀ values between about 0.001 toabout 10 micromolar micromolar based on inhibiting a contraction of arat bladder muscle strip in vitro; even more preferably the Myosin IIATPase inhibitor compounds of formula 1 and formula II in their activeform have an IC₅₀ values between about 0.001 to about 5 micromolarmicromolar based on inhibiting a contraction of a rat bladder musclestrip in vitro. Most preferred Myosin II ATPase inhibitor compounds offormula 1 and formula II in their active form have a rat bladder striptest IC₅₀ values between about 0.001 to about 1.0 micromolar micromolarbased on inhibiting a contraction of a rat bladder muscle strip invitro. The rat bladder muscle strip test can also be useful for testinga conversion of a prodrug of formula I or a prodrug of formula II “toits active form”. In addition, the rat bladder strip IC₅₀ test can alsobe used to evaluate or discover a compound's light stability orinstability or assess the the potency of a metabolite of a compound ofthe invention as an inhibitor of Myosin II ATPase or as having someother effect, as well as to help discover other chemical changes of acompound of the invention that might alter its rat bladder muscle striptest IC₅₀ value.

Provided is a method of treating an overactive bladder in a patient, themethod comprising administering to the patient an effective amount of aMyosin II ATPase inhibitor compound, useful in its active form,comprising a3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-oneas represented by structural Formula 1, or a pharmaceutically-acceptablesalt, racemic mixture, enantiomer, or prodrug thereof,

wherein X is selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,(C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent W,the substituent W selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent Q, the substituent Q selected from thegroup consisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, napthyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; wherein Y is selected from thegroup consisting of hydrogen, methyl, hydroxy, fluoro, chloro, bromo,iodo, nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂ (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a carbon atom with a substituent Z, thesubstituent Z selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent T, the substituent T selected from thegroup consisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, (C₄₋₁₀)aryl, napthyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; and wherein said Myosin IIATPase inhibitor compounds of Formula 1, have an IC₅₀ value betweenabout 0.001 to about 50 micromolar based on a test of an inhibition ofcontraction of a rat bladder muscle strip in vitro.

For the present invention, the meaning of a (C₁₋₆)alkyl includes methyl,ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl,pentyl, 2-methylbutyl, isopentyl, tert-pentyl, neopentyl, hexyl,1-methyl-pentyl, 2-methylpentyl, 3-methylpentyl, isohexyl, tert-hexyl,neohexyl, and 2,2-dimethyl-butyl.

For the present invention, the meaning of a (C₂₋₆)alkenyl includes (a) a(C₂₋₆)alkyl minus 2 hydrogens and having 1 carbon double bond; (b) a(C₂₋₆)alkyl missing 4 hydrogens and having 2, carbon double bonds; and(c) (C₂₋₆)alkyl missing 6 hydrogens and having 3 carbon double bonds.For the present invention, the meaning of a (C₂₋₆)alkynyl includes a(C₂₋₆)alkyl minus 4 hydrogens and having one triple carbon bond.

For the present invention, the meaning of a (C₁₋₆)haloalkyl includes a(C₁₋₆)alkyl having one or a plurality of halogen atoms (F, Cl, Br, or I)substituents in place of a hydrogen atom on a carbon atom of the alkylgroup.

For the present invention, the meaning of a (C₁₋₆)alkoxy includesmethoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, isobutoxy,tert-butoxy, pentoxy, 2-methylbutoxy, isopentoxy, tert-pentoxy,neopentoxy, hexoxy, 1-methylpentoxy, 2-methylpentoxy, 3-methylpentoxy,isohexoxy, tert-hexoxy, neohexoxy, or 2,2-dimethylbutoxy.

For the present invention, the ring group may be fully saturated withhydrogen or have 1-3 double bonds or maximum aromatic ring character asone of skill in organic chemistry would appreciate or understand for anyof the fore-mentioned optional substituents based definitive rules fornomenclature of organic chemistry as taught by IUPAC (the InternationalUnion of Physics and Chemistry 1957 Rules pages C1-C-73 in the CRC PressHandbook of Chemsistry and Physics, 54th Edition, Cleveland, Ohio)supplemented by the textbook “Organic Chemistry”, 6th Edition, Morrison& Boyd, 1992.

For the present invention, a (C₃₋₁₂)cycloalkyl means a cyclic alkyl ringof 3 to 12 carbons. Examples include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl,cyclodecyl.cycloundecyl, and cyclododecyl.

For the present invention, the meaning of a (C₄₋₁₂)cycloalkenyl means aring with 4 to 12 carbon atoms and includes a double bond or a pluralityof double bonds in the ring and includes all contemplated isomers ofcyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,cyclononenyl, cyclodecenyl.cycloundecenyl, and cyclododecenyl rings.

For the present invention, the meaning of a (C₄₋₁₂)heterocyclic includesa non-aromatic ring system having 4 to 12 atoms in one ring, or in twofused rings, said ring or rings having in total at least one or morecarbon atoms replaced by one or more heteroatoms selected from the groupconsisting of N, O, and S. For the present invention the meaning of a(C₄₋₁₂)heterocyclic includes rings such as for example, axiridinyl,oxiranyl, thiiranyl, oxaziridinyl, dioxiranyl, azetidinyl, oxetanyl,thietanyl, diazetanyl, dioxzetanyl, dithietanyl, pyrrolidinyl,tetrahydrofuranyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,isoxalidinyl, thiazolidinyl, isothiazolidinyl, dioxolanyl, dithiolanyl,piperidinyl, oxanyl, thianyl, piperazinyl, morpholinyl, thiomorpholinyl,dioxanyl, or dithianyl ring. Some (C₄₋₁₂)heterocyclic ring examplesnamed to indicate where a bond may be attached include:3-tetrahydrofuranyl, 2-tetrahydropyranyl, 3-tetrahydropyranyl,4-tetrahydropyranyl, [1,3]-dioxalanyl, [1,3]-dithiolanyl,[1,3]-dioxanyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl,2-morpholinyl, 3-morpholinyl, 4-morpholinyl, 2-thiomorpholinyl,3-thiomorpholinyl, 4-thiomorpholinyl, 1-pyrrolidinyl, 2-pyrrolidinyl,3-pyrorolidinyl, 1-piperazinyl, 2-piperazinyl, 1-piperidinyl,2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl, 5diazolonyl, N-substituted diazolonyl, and 1-pthalimidinyl.

For the present invention, the meaning of a (C₄₋₁₂)aryl includes acarbocyclic aromatic ring or two rings spiro or fused having 4 to 12carbon atoms and significant aromatic character. (C₄₋₁₀)Aryl examplesinclude cyclobutadiene, cyclopentadiene, pentalenyl, phenyl, indenyl,azulenyl, and naphthyl. Aromatic character in such rings becomesapparent when there are two or more double bonds in each ring.

For the present invention, the meaning of a (C₄₋₁₂)heteroaryl includesone aromatic ring or two aromatic rings arranged in some way or in twofused rings, said ring or rings having in total 4-12 atoms, of which3-11 are carbon atoms, wherein at least one or more carbon atoms isreplaced by a heteroatom selected from the group consisting of N, O, andS. (C₄₋₁₂)heteroaryl examples include imidazolyl, isoimidazolyl,thienyl, furanyl, pyridyl, pyrimidyl, pyranyl, pyrazolyl, pyrrolyl,pyrazinyl, thiazolyl, isothiazolyl, oxazolyl, isooxazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, and tetrazolyl. benzofuranyl,isobenzofuranyl, pyrrolyl, indolyl, isoindolyl, thiophenyl,benzothiophenyl, benzo[b]thienyl, benzo[c]thiophenyl, imidazoyl,oxazolyl, benzoxazoyl, isoxazoyl, benzisoxazoyl, thiazoyl,benzothiazolyl, pyridinyl, quinolinyl, isoquinolinyl, pyrazinyl,quinoxalinyl, acridinyl, pyrimidinyl, and cinnolinyl.

(S)-blebbistatin, (−)-blebbistatin, (−)-(S)-blebbistatin, andracemic-blebbistatin are some of the preferred compounds of the presentinvention. A racemic mixture of blebbistatin enantiomers has usefulpotent Myosin II ATPase inhibitory activity. An (S)-enantiomer or an(−)-enantiomer of blebbistatin or another compound of the invention canbe a potent Myosin II ATPase inhibitor also as a racemic mixture withits corresponding (R)-enantiomer or (+)-enantiomer.

(R)-blebbistatin has a much higher IC50 value than (S)-blebbistatin.Thus (R)-blebbistatin can be used as a “control” during in vitro bladdrsmooth muscle contraction studies of the inhibitory effects of(S)-blebbistatin or racemic blebbistatin.

For the present invention note that blebbistatin can be named in avariety of ways. The names (−)-blebbistatin, (S)-blebbistatin and(S)-(−)-blebbistatin refer to the same enantiomer of the same compound(Lucas-Lopez, C., Patterson, S, Blum, T., Straight, A. F., Toth, J.,Slawin, A. M. Z., Mitchison, T. J., Sellers and, J. R., Westwood, N. J.“Absolute Stereochemical Assignment and Fluorescence Tuning of the SmallMolecule Tool, (−)-Blebbistatin” European J. Organic Chemistry. (2005),pp 1736-1740.). IUPAC (International Union of Physicists and Chemists)publications state a preference in chemical naming of a compound thatthe chemical name list its group substituents in an alphabetic orderwith the core ring system last named if the ring system is selected asthe core molecule from which there are the group substituents.Accordingly, (S)-blebbistsatin is named(S)-3aS-hydroxy-6-methyl-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-one).Also accordingly then (−)-blebbistatin is named(−)-3aS-hydroxy-6-methyl-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-one).However (S)-blebbistatin might be named(S)-6-methyl-1-phenyl-1,2,3,3a-tetrahydro-3aS-hydroxy-4H-pyrollo[2,3b]quinolin-4-one).Also (−)-blebbistatin might be named(−)-6-methyl-1-phenyl-1,2,3,3a-tetrahydro-3aS-hydroxy-4H-pyrollo[2,3b]quinolin-4-one).For the present invention, these names for the (−) enantiomer ofblebbistatin should be considered to mean the same enantiomer ofblebbistatin.

Synthesis of Formula 1 and Formula II compounds of the present inventioncan use intermediate organic chemical compounds and chemical reagents,the later reagents are easily purchased. The substituted intermediateorganic chemical compounds such as the substituted anthranilic acidesters intermediate compounds, can now be bought as well or custom madeby some chemical companies and purchased for making a compound of thepresent invention. Said intermediate organic chemical compounds can bemade or bought and optionally may be used in a form having a chemicalblocking groups (“protecting groups”) that protects some of thecompound's group substituents during a synthetic chemical reaction.

When buying an intermediate chemical compound for the present invention,there are several approaches to select from. One approach is to use thechemical database and pharmaceutical building block resources atMaybridge.com to obtain the internmediate compound. A second approach isto use MolPort.com, a website resource that enables a centralizedordering of chemical compounds from over 200 chemical companies andprovides a database of commercially available chemical compounds. Athird approach is to contact and then buy the intermediate chemicalcompounds and fine reagents from the Sigma-Aldrich Chemical Company (St.Louis, Mo., U.S.), Molport Chemical Company (Riga, Latvia), orThermo-Fisher-Scientific subsidiary Maybridge Chemical Company in theU.S. or in Tintigal, U.K. The compound or its intermediate may besynthetically prepared in a custom batch order by contacting MaybridgeChemical Company service CustomBlocks™ (link is maybridge.sales atThermofisher.com) a subsidiary of Thermo Fisher Scientific Inc. Thethird approach which is working with Maybridge Chemical Company is apreferred means for obtaining complex intermediates of a compound of theinvention.

A compound of formula I or formula II of the present invention or forthat matter any chemical structure for a compound that inhibits or mayconceivably inhibit Myosin II ATPase, can be written out on paper as amolecular structure. Optionally said molecular structure can be sent toa chemical company such as Maybridge Chemical Company for its customsynthesis for a fee. Thus, it is contemplated for the present invention,that a compound of the invention that may be a Myosin II ATPaseinhibitor can be conceived, and then can be custom-made by a chemicalcompany as a means for enabling one of skill in the art to practice anyaspect of the present invention as needed.

One of ordinary skill in the art of organic chemical synthesis willappreciate that the synthesis of the Myosin II ATPase compounds of thepresent invention is facilitated when performed by an experienced Ph.D.organic chemist having a resourceful, careful, and safety-conscious setof skills and who has been working in medicinal drug discovery at apharmaceutical company or university for several years in heterocyclicorganic chemicals synthesis. Said chemist will have experience inpurchasing and effectively using various intermediate organic compoundsfrom chemical supply companies. Said chemist will select a specific(−)-3aS-hydroxy-quinolone compound 5 to be made. Then, using a disclosedsynthetic route of the present invention, said chemist will know eachspecific chemical compound intermediate in the synthetic route makingthe formula I and formula II compounds of the present invention.

To one of skill in organic synthesis, some synthetic routes to compoundsof the present invention will be more efficiently performed using“protecting groups” (also known as “blocking groups”) on theintermediate compounds. Protecting groups are well known in the art oforganic chemical synthesis as a means for increasing the selectivity ofan organic chemical reaction. Selective chemical reactions are known toenable a higher purity and higher yield reaction product, advantageouslyfacilitating subsequent reactions. It is an object of the presentinvention that synthesis of the compounds of formula I and formula IIare well-controlled so that only the unprotected chemical groupsubstitutents in the intermediates react in each reaction step. Analcohol, amine, carbonyl, carboxylic, phosphate, or terminal alkynegroups may need to be chemically-blocked before a reaction step to keepthem from participating in the reaction.

Protection of alcohols can be done using a blocking group for protectionof alcohol groups selected from the group consisting of acetyl (Ac),benzoyl (Bz), benzyl (Bn, Bnl), β-methoxyethoxymethyl ether (MEM),dimethoxytrityl, [bis-(4-methoxyphenyl)phenylmethyl] (DMT),methoxymethyl ether (MOM),methoxytrityl[(4-methoxyphenyl)diphenylmethyl, MMT), p-methoxybenzylether (PMB), methylthiomethyl ether, pivaloyl (Piv), tetrahydropyranyl(THP), Trityl (triphenylmethyl, Tr), Silyl ether, trimethylsilyl ether(TMS), tert-butyldimethylsilyl ether (TBDMS),tri-iso-propylsilyloxymethyl ether (TOM), triisopropylsilyl ether(TIPS), methyl ethers (ME), and ethoxethyl ethers (EE).

The alcohol protecting groups are removed in different ways. An acetylgroup is removed by acid or base. A benzoyl group is removed by acid orbase, more stable than Ac group. A benzyl group is removed byhydrogenolysis. A β-Methoxyethoxymethyl ether group is removed by acid.A dimethoxytrityl group is removed by weak acid. A methoxymethyl ethergroup is removed by acid. A methoxytrityl group is removed by acid andhydrogenolysis. A p-methoxybenzyl ether group is removed by acid,hydrogenolysis, or oxidation. A methylthiomethyl ether group is removedby acid. A pivaloyl group is removed by acid, base or reducing agents. Apivalyl group is substantially more stable than other acyl protectinggroups. A tetrahydropyranyl group is removed by acid. Trityl is removedby acid and hydrogenolysis. A silyl ether group such as TMS, TBDMS, TOM,and TIPS, are removed by acid or fluoride ion (such as NaF, TBAF(Tetra-n-butylammonium fluoride, HF-Py, or HF-NEt3)). A methyl ethergroup is cleaved by TMSI in dichloromethane, acetonitrile, orchloroform; alternatively cleave silyl ethers using BBr3 inDichloromethane. An ethoxyethyl ethers group can be cleaved in 1N HCl.

Protection of amines can be done using a blocking group for protectionof amine groups selected from the group consisting of (1) carbobenzyloxy(Cbz), (2) p-methoxybenzyl carbonyl (Moz or MeOZ), (3)tert-butyloxycarbonyl (BOC), (4) 9-fluorenylmethyloxycarbonyl (FMOC),(5) acetyl (Ac), (6) benzoyl (Bz), (7) benzyl (Bn), (8) carbamate, (9)p-methoxybenzyl (PMB), (10) 3,4-dimethoxybenzyl (DMPM), (11)p-methoxyphenyl (PMP), (12) tosyl (Ts) and other sulfonamides (Nosyl &Nps).

The amine protecting groups are removed in different ways. Acarbobenzyloxy group is removed by hydrogenolysis. A p-methoxybenzylcarbonyl group is removed by hydrogenolysis. It is more labile than aCbz group. A tert-butyloxycarbonyl group is removed by concentrated,strong acid. (such as HCl or CF₃COOH). A 9 fluorenylmethyloxycarbonylgroup is removed by a base, such as piperidine. An acetyl group isremoved by treatment with a base, most often, with aqueous or gaseousammonia or methylamine. An acetyl group is too stable to be readilyremoved from aliphatic amides. A benzoyl group is removed by treatmentwith a base, most often with aqueous or gaseous ammonia or methylamine.A benzoyl like an acetyl group is too stable to be readily removed fromaliphatic amides. A benzyl group is removed by hydrogenolysis. Acarbamate group is removed by acid and mild heating. A p-methoxybenzylgroup is removed by hydrogenolysis, and is more labile than a benzylgroup. A 3,4-dimethoxybenzyl group is removed by hydrogenolysis, and ismore labile than a p-methoxybenzyl group. A p-methoxyphenyl group isremoved by Ammonium cerium(IV) nitrate. A tosyl group is removed byconcentrated acid such as HBr or H₂SO₄; or by a strong reducing agentsuch as sodium in liquid ammonia or sodium naphthalenide. Othersulfonamides such as Nosyl and Nps can be removed by samarium iodide ortributyltin hydride.

Protection of carbonyl groups can be done using a carbonyl blockinggroup selected from the group consisting of (1) acetals and ketals(which are removed by acid and cleavage of an acyclic acetals is easierthan a cleavage of a cyclic acetal), (2) acylals (which are removed by aLewis acid), and (3) dithianes which are removed by metal salts oroxidizing agents.

Protection of carboxylic acid groups can be done using a carboxylblocking group selected from the group consisting of (1) methyl esters(which are removed by acid or base), (2) benzyl esters (which areremoved by hydrogenolysis), (3) tent-butyl esters (which are removed byacid, base and some reducing agents), (4) silyl esters (which areremoved by acid, base and organometallic reagents), (5) orthoesters(which are removed by mild aqueous acid to form an ester, said esterthen removed with a facility depending on its ester properties), and (6)oxazoline (which is removed by strong hot acid with a pH<1 and at atemperature above 100° C. or which is removed by alkali creating pHgreater than 12 and at a temperature above 100° C.), but said blockinggroup does not protect against LiAlH₄, organolithium reagents orGrignard (organomagnesium) reagents.

Protection of phosphate groups can be done using a phosphate blockinggroup selected from the group consisting of (1) 2-cyanoethyl (which isremoved by mild base), and (2) methyl (which removed by a strongnucleophiles such as a mixture of thiophenol with triethanolamine).

Protection of a terminal alkyn group can be done using a terminal alkynblocking group selected from the group consisting of (1) propargylalcohols in the Favorskii reaction, and (2) silyl groups, especially inprotection of the acetylene itself.

For example a chemist can select a specific (−)-3aS-hydroxy-quinolonecompound 5 of the present invention to be make. Based on knowledge aboutthe use of protecting groups and using a disclosed synthetic route ofthe present invention, a chemist can make reasonable expectations as towhat each chemical intermediate will need to be in the synthetic routethat makes the formula I and formula II compounds of the presentinvention. For example, the methyl anthranilate intermediate compound 2(depicted below in a reaction scheme diagram) has an X substitutent atthe five position and a Y substitutent at the six position. When the Xand the Y substitutent groups are free of any “protecting group”, thenthese X and Y correspond to the X and Y disclosed in Formula I andFormula II compounds of the present invention. Thus, below theintermediate compound 2 which is a methyl anthranilate (2-amino-benzoicacid methyl ester) is chosen to provide specific X and Y substituents.As mentioned said intermediate compound 2 may be bought, made by thechemist, or bought custom-made from a company.

For example a synthesis route for a Formula 1 compound comprises makingthe corresponding intermediate amidine compound 3 by a Step (a) process.Phosphorus oxychloride (POCl₃, 1.0 mmol (millimole) is added, drop-wise,to a solution of N-phenyl-2-pyrollidinone 1 (1.1 mmol) in drydichloromethane (5.0-15 ml [milliliters]) with stirring for 3 hours atroom temperature (22° C.). A solution of 1.1 mmol anthranilate 2 in drydichloromethane (15-60 ml) then is added. The mixture is refluxed for16-80 hours, is cooled and then is concentrated under vacuum. The solidis dissolved in 0.30 N hydrochloric acid (100 ml) and is extracted withdicloromethane (3×100 ml). Then to the aqueous phase is added 100 mlethyl acetate and slowly using aqueous 2 molar sodium hydroxide, the pHis raised to pH 8.0. The aqueous phase is further extracted with ethylacetate (3×100 ml). All ethyl acetate extracts are combined and driedwith anhydrous MgSO₄. The dried ethyl acetate extracts which hold theproduct as a base, are dried by vacuum to yield amidine solid 3. Theprocess of a Step (a) reaction scheme is below depicted:

For example, a synthesis route for a Formula 1 compound comprises makingthe corresponding intermediate quinolone compound 4 by a Step (b)process. A reaction flask holding a solution of the amidine 3 (1.0 mmol)in 50 ml anhydrous THF (tetrahydrofuran is cooled to −78° C. (reactionflask is cooled in acetone/dry ice mix) and is stirred for 15 minutes. Asolution of lithium bis(trimethylsilyl)amide (LiHMDS) in THF (1 molar,3.0 mmol) is added drop-wise to the amidine in THF solution to react.The reaction mixture is allowed to warm to 0° C. over 3 hours or allowedto warm to 22° C. (room temp.) over 12 hours. The reaction is thenquenched with saturated aqueous ammonium chloride (150 ml) and theaqueous phase is extracted with dichloromethane (4×100 ml), the CH₂Cl₂extracts are combined and are dried using anhydrous magnesium sulfate.After filtering, the anhydrous extract is concentrated by vacuum. Thequinolone 4 is purified by flash column chromatography on silica geleluting with (50-100% ethyl acetate—PE 40-60 [wherein PE 40-60 is thefraction of light petroleum ether boiling in the range of 40-60° C.]).The process of a Step (b) reaction scheme is below depicted.

For example, a synthesis route for a (−) enantiomer Formula 1 compound:Step (c1) process forms a is (−)-3aS-hydroxy-quinolone compound 5synthesis. Quinolone 4 (1.0 mmol) in dry THF (10-50 ml) as a solution isadded drop-wise to lithium bis(trimethylsilyl)amide (LiHMDS) in THF (1.0molar, 1.2 equiv.) in dry THF (2.0-7.0 ml) at −78° C. (reaction flask iscooled in acetone/dry ice mix) under argon. Mixture is stirred (for 30min.) at −78° C. In dry THF 4-12 ml), 2.4 mmoles of(−)-(8,8-dichlorocamphorylsulfonyl)oxaziridine [Davis oxaziridinemethod] in solution is added via a cannula. After 16 hours reactiontime-mixing at −10° C., a saturated aqueous ammonium iodide solution(5.0 ml, 10 mmol) and diethyl ether (5.0-10 ml) are added. Then themixture is extracted with diethyl ether (3×10 ml). Combined organicextracts are dried with magnesium sulfate, filtered and concentratedusing a vacuum. The concentrate is added to 100 ml dichloro-methane and100 ml of hydrochloric acid (0.3 molar). The product is now a salt inthe aqueous phase portion. The aqueous phase is isolated and washed withCH₂Cl₂ (3×100 ml). Then the aqueous extract is slowly alkalinized, withaqueous 2.0 molar sodium hydroxide until the pH is 8.0. The product, nowa base, is suspended in the aqueous phase, said aqueous phase thenextracted with ethyl acetate (3×100 ml). The combined ethyl acetateextracts are dried with magnesium sulfate, filtered and concentrated toobtain the base product which is substantially the (−) enantiomer,namely, (−)-3aS-hydroxy-quinolone compound 5. For the present inventionit is also termed the (−)-X,Y-substituted Formula 1 compound and termedthe(−)3aS-hydroxy-1-phenyl-X-Y-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-one).Step (c1) reaction scheme is depicted below.

For example, optionally, a (−)-3aS-hydroxy-quinolone compound 5 can berecrystallized in acetonitrile or a similar solvent, using standardrecrystallization methods of one of skill in the art, to increase itsenantiomeric purity from below 90% E.E. to about 99.5% E.E. as needed.The E.E. % values are determined using chiral HPLC. In addition, acompound of the present invention will exist in different polymorphforms. Polymorphs can have advantages such as improved solubility,improved chemical stability, and improve a characteristic of or theprocess of making a pharmaceutical formulation. The present inventionincludes any polymorphs of intermediates, and of compounds of formula Iand formula II.

For example, using above described synthetic routes for making 1millimole of (−)-blebbistatin and starting with 5-methyl anthranilatemethyl ester 1 (X═CH₃, Y═H), a 41% yield has been reported for theformation of the corresponding amidine 3; a 90% yield has been reportedfor the cyclization of amidine 3 to quinolone 4; an 82% yield has beenreported for 3a-hydroxylation of quinolone 4 to (−)-3a-hydroxy-quinolone5, said (−)-3a-hydroxy-quinolone 5 compound has been reported to have anenantiomeric excess of 82% (E.E. 82%); and following a singlecrystallization in a solvent such as acetonitrile, said(−)-3a-hydroxy-quinolone 5 compound has been reported to have an opticalpurity of 99.5% E.E. [(1) Lucas-Lopez, C., Allingham, J S., Lebl, T.,Lawson, C. P. A. T., Brenk, R., Sellers, J. R., Rayment, I., Westwood,N. J. “The small molecule tool (S)-(−)-blebbistatin: novel insights ofrelevance to myosin inhibitor design” Organic & Biomolecular Chemistry(2008); vol. 6:pp 2076-2084. (2) Lucas-Lopez, C., Patterson, S, Blum,T., Straight, A. F., Toth, J., Slawin, A. M. Z., Mitchison, T. J.,Sellers and, J. R., Westwood, N. J. “Absolute Stereochemical Assignmentand Fluorescence Tuning of the Small Molecule Tool, (−)-Blebbistatin”European J. Organic Chemistry. (2005), pp 1736-1740. (3) Patterson, S.,Lucas-Lopez, C., Westwood, N. J. “Selective Chemical Interventions inBiological Systems: The Small Molecule Tool, (S)-(−)-Blebbistatin” pp147-166, Beilstein-Institute, The Chemical Theatre of BiologicalSystems, (May 24th-28th, 2004) Bozen, Italy.]

For example, a synthesis of (±) racemic Formula 1 compounds:(−)-3aS-hydroxy-quinolone compound 5 and (+)-3aR-hydroxy-quinolonecompound 6 from racemic hydroxylation of carbon 3a in quinolone compound4 is shown in Step (c2) below. Quinolone compound 4 is racemicallyhydroxylated at carbon 3a when dissolved in dimethylsulfoxide (DMSO) orin THF and the solution is exposed to air. The rate and yield of theracemic 3a carbon hydroxylation of quinolone compound 4 is increased byirradiation of said solution of compound 4 by a medium pressure mercurylamp (400 watt, or upon irradiation at 368 nanometers of the compound onsilica gel. A 3 hour exposure to the mercury lamp at 25° C., gives ayield of 26% in DMSO and a yield of 29% in THF for making a racemicmixture of the 3aR- and 3aS-hydroxy-quinolone compound 5 (specifically,(±)-blebbistatin, has been reported (Lucas-Lopez, C., Patterson, S,Blum, T., Straight, A. F., Toth, J., Slawin, A. M. Z., Mitchison, T. J.,Sellers and, J. R., Westwood, N. J. “Absolute Stereochemical Assignmentand Fluorescence Tuning of the Small Molecule Tool, (−)-Blebbistatin”European J. Organic Chemistry. (2005), pp 1736-1740.). Step (c2) racemicreaction scheme is depicted below which forms both hydroxyl 3aenantiomers from quinolone intermediate compound

A third embodiment of the invention provides a method of treating anoveractive bladder in a patient, the method comprising administering tothe patient an effective amount of a Myosin II ATPase inhibitorcompound, useful in its active form, comprising a substituted3a-hydroxy-1-phenyl-1,2,3,3a-tetrahydro-4H-pyrollo[2,3b]quinolin-4-oneas represented by structural Formula II, or apharmaceutically-acceptable salt, racemic mixture, enantiomer, orprodrug thereof,

wherein X is selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, optionally substituted at a carbon atom with a substituent W,the substituent W selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂ (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, acetamido, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl,asparginyl, glutamyl, lysinyl, histidinyl, arginyl, aspartyl, andglutamyl, and optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent Q, the substituent Q selectedfrom the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic,(C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; wherein Y isselected from the group consisting of hydrogen, methyl, hydroxy, fluoro,chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,optionally substituted at a carbon atom with a substituent Z, thesubstituent Z selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyklalanyl, tyrosinyl, trytophanyl, threoninyl, cystinyl, asparginyl,glutamyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andoptionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent T, the substituent T selected from thegroup consisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, (C₄₋₁₀)aryl, napthyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; and wherein an atom from X isproximal to an atom of Y and said atoms form a chemical bond; saidchemical bond forms a ring from X and Y; wherein the ring is anortho-fused to an existing ring of Formula II having X and Y.

The invention also provides pharmaceutical compositions comprising aneffective amount of a Myosin II ATPase inhibitor compound, useful in itsactive form, as a method of treating an overactive bladder in a patient.The compounds of the invention comprise optionally substituted compoundsand their pharmaceutically-acceptable salts; saidpharmaceutically-acceptable salts are well known in the art. Compoundsof the present invention may be formulated as salts using an acid toimprove the aqueous solubility of the compound. Compounds of the presentinvention are ionized in acidic solutions which ionized compound is morewater soluble. Suitable pharmaceutical salts of the compounds of thepresent invention include the hydrochloride, sulfate, acetate,phosphate, maleate, citrate, nitrate, tosylate, mesylate and besylatesalts. Preferably the salt is a chloride, acetate, sulfate, nitrate, orcitrate or besylate salt; or most preferably a chloride salt. There is atertiary amine at the 1 position in the pyrrole ring and at the 1position of the quinolin-4-one ring in Formula 1. Thus at least 1 to 2or more molar equivalents of an acid are needed to protonate the basicFormula 1 compound to form its salt forms. Prodrug formulations of theinvention include esters of the Formula I and II compounds that areformed by reacting a 3a-hydroxyl group of the formual I or Formula IIcompound with acetic acid, benzoic acid or another suitable carboxylicacid to make the corresponding acetate or benzoate or other suitablecarboxylic acid ester. The 3a-hydroxy group of a compound of formula Iand of formula II of the present invention is essential for the compoundto be in “its active form”. Compounds of Formula I and II with achemically modified group such as an ester instead of a hydroxyl atposition 3a will be not be “in an active form” as a Myosin II ATPaseinhibitor of the invention.

One of skill in the art of organic chemical molecule synthesis methodscan use known blocking groups, nucleophilic substitution reactions andaddition organic chemistry reactions in various combinations tosynthesize the substituted (S)-(−)-blebbistatins of the presentinvention. Useful synthesis procedures and guidances for the synthesisof racemic and enantiomeric compounds of the present invention may befound also in the following articles: (1) Lucas-Lopez, C., Allingham, J.S., Lebl, T., Lawson, C. P. A. T., Brenk, R., Sellers, J. R., Rayment,I., Westwood, N. J. “The small molecule tool (S)-(−)-blebbistatin: novelinsights of relevance to myosin inhibitor design” Org. Biomol. Chem.2008; 6:2076-2084; (2) Patterson, S., Lucas-Lopez, C., Westwood, N. J.“Selective Chemical Interventions in Biological Systems: The SmallMolecule Tool, (S)-(−)-Blebbistatin” pp 147-166, Beilstein-Institute,The Chemical Theatre of Biological Systems, May 24TH,-28TH, Bozen,Italy; and (3) Lucas-Lopez, C, Patterson, S., Blum, T., Straight, A. F.,Toth, J., Slawin, A. M. Z., Mitchison, T. J., Sellersand, J. R.,Westwood, N. J. “Absolute Stereochemical Assignment and FluorescenceTuning of the Small Molecule Tool, (−)-Blebbistatin” Eur. J. Org. Chem.2005, 1736-1740.

For the present invention, many routes of administering of a compound ofthe present invention are contemplated as a method of treating anoveractive bladder of a patient. The drug may be in the form of a base,or a pharmaceutically acceptable salt. The drug may be formulated withconventional excipients whose selection is within ordinary skill in theart of using pharmaceutical excipients to make a pharmaceuticalformulation. The design of a formulation is well-known in the art to bemodified to accommodate a particular route of administration. Routes ofadministration for the present invention shall include conventional andnon-conventional routes. A preferred route of administration of a MyosinII ATPase inhibitor compound minimizes subjecting a patient beingtreated for OAB from possible systemic side effects of the compound. Apreferred route of administration is direct administration of a MyosinII ATPase inhibitor to the overactive bladder of the patient. Theinhibitor compound may be vesicularly administered by any suitable meanssuch as by a cannula or a pump.

Pharmaceutical compositions of the present invention may contain up to85 wt % of a compound of the invention. More typically, thepharmaceutical composition contains up to 50% weight of a compound ofthe invention. Preferred pharmaceutical compositions are sterile andpyrogen free. Preferably the pharmaceutical compositions provided by theinvention contain a compound of the invention which is a substantiallycompound such as a pure (−) optical isomer, a racemic mixture, or aprodrug of a compound of the invention. A therapeutically effectiveamount of a compound of the invention, its racemic mixture, anenantiomer, or prodrug thereof, that is administered to an OAB patientis about 0.01 mgs to about 50 mg per kg of body weight, depending uponthe activity of the compound. Some other factors may guide in selectingthe pharmaceutical dose, including the age, weight and conditions of thepatient to be treated, the type and severity of the disease and thefrequency and route of administration.

Solutions for injection or infusion may contain as carrier, for example,sterile water or preferably they may be in the form of sterile, aqueous,isotonic saline solutions. Preferably a dose of a compound of theinvention is administered intervesicularly. Intervesicularadministration means administration into the bladder where urine isstored, said urine storage space known as intervesicular space of thebladder.

When for example, a dose of a compound of the invention is administeredintervesicularly, then the IC₅₀ of the compound in its active form canbe useful number to use in estimating a therapeutically effective dose(amount) of the compound to be administered as a treatment of anoveractive bladder of a patient, said IC₅₀ determined from (a) an invitro study of the inhibition of adult rat bladder Myosin II ATPaseactivity or (b) an in vitro study of the inhibition of a human bladderMysin II ATPase activity or (c) an in vitro study of the inhibition of aadult rat bladder muscle strip contraction. The IC₅₀ of the compound inits active form determined from the in vitro study of the inhibition ofa adult rat bladder muscle strip contraction is described in detail inthe written description of the present invention.

An effective intervesicular dose can be achieved if the intervesicularurine concentration of the compound of the invention is adjusted tobetween about 0.1 IC₅₀ to about 3 IC₅₀ of the compound, preferably about0.5 to about 2.0 IC₅₀, of the compound more preferably about 0.80 toabout 1.5 IC₅₀. of the compound. Most preferably the intervesicularurinary concentration of the compound is equivalent to about 1.0 IC₅₀ ofthe compound of the invention, and if the IC₅₀ of the compound is 1micromolar to 20 to micromolar, then the desired urinary concentrationof the compound in its active form would be about 1 to 10 micromolar. Apharmaceutical solution for intervesicular instillation can be designedaccordingly, optionally including an adjustment of the dose assuming adilution effect of the dose by the urine in the bladder.

The IC₅₀ of compounds of the invention are typically between 0.0001micromolar to 50 micromolar and accordingly for said compounds, then thedesired urinary concentration of the compound in its active form wouldbe between about 0.0001 micromolar to about 50 micromolar. Compoundshaving an IC₅₀ greater than 50 micromolar may be too impotent andnon-selective whereas compounds having an IC₅₀ less than 0.0001 maydisadvantageously function as an irreversible inhibitor as ifcovalently-linked irreversibly to the Myosin ATPase. Optionally, themethod of treating on overactive bladder of a patient includes betweenabout 0.01 to about 3 IC₅₀ of a phosphodiesterase five inhibitorcompound, preferably a PDE5 inhibitor such a sildenafil citrate,tadalafil or vardenafil. The IC₅₀ of the PDE5 inhibitor may bedetermined by in vitro test methods known in the art or from publishedvalues for the PDE5 inhibitor or from the FDA approved package insert ofthe available PDE5 inhibitor.

For oral or parenteral administration a compound of the invention may beformulated for administration with a pharmaceutically acceptable carrieror diluent For example, solid oral forms may contain, together with theactive compound, diluents, e.g. lactose, dextrose, saccharose,cellulose, corn starch or potato starch lubricants, e.g. silica, talc,stearic acid, magnesium or calcium stearate, and/or polyethyleneglycols; binding agents; e.g. starches, arabic gums, gelatin,methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone;disaggregating agents, e.g. starch, alginic acid, alginates or sodiumstarch glycolate; effervescing mixtures; dyestuffs; sweeteners; wettingagents, such as lecithin, polysorbates, laurylsulphates; and, ingeneral, non toxic and pharmacologically inactive substances used inpharmaceutical formulations. Such pharmaceutical preparations may bemanufactured in known manner, for example, by means of mixing,granulating, tableting, sugar coating, or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions andsuspensions. The syrups may contain as carriers, for example, saccharoseor saccharose with glycerine and/or mannitol and/or sorbitol.

Light-sensitive compounds of the invention, such as blebbistatin, mustbe shielded from light. Said compounds in pharmaceutical formulationswill have light protection barriers, for example a suitable dye andopacifier excipient in oral formulations, or light-shielding storage andadministration containers for a parenteral, i.v., or intervesicularformulation. Optionally a compound of the invention is substituted atthe 7 position, if a formula 1 with a nitro group to reduce lightsensitivity which causes the compound to become less active orinactivated.

Suspensions and emulsions may contain as carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections may contain, together with theactive compound, a pharmaceutically acceptable carrier, e.g. sterilewater, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and ifdesired, a suitable amount of lidocaine hydrochloride.

For the present invention, a preferred route of administration of themedication to treat OAB is a direct route to minimize side effects ofthe medication. Medication for the present invention is a drug ormixture of drugs comprising a compound of the present invention, thecompound may be for example blebbistatin or an “other drug” as “otherdrug” is defined for the present invention. “Other drug” is a group ofchemical substances or drugs which include a smooth muscle relaxantknown or to be discovered or a prodrug of a metabolite.

Preferably the medication is deposited into the intravesical space ofthe bladder (the chamber of the chamber) and remains there a long timeso that the treatment of a patient's OAB condition does not requirefrequent drug administration to the patient. For the present invention,formulations of the medication in the bladder may exist in a reservoirof a small pump (“a drug storing-pumping device”) that slowly deliversthe drug into the bladder, the pump and reservoir subsiding inside thebladder itself. The device may be placed inside the bladder by asurgical means, by a syringe, or via a cannula. Slow-release formulationpreferably would be delivered via the ureter. Advantageously, placementof the drug into the bladder reduces its metabolism, systemic sideeffects, requires a lower dose, provides faster onset and higher druglevels in the urine (optimizes pharmacokinetics). Another preferableroute of administration via the ureter comprises a liquid formulation, asuspension or formulation capable of forming a viscous gel with orwithout slow time release pellets. Such administration is readilyaccomplished by a pressure injection involving a cannula in the ureteror urethra, or by pressing a blunted syringe cannula tip into a ureteror urethra so that a non-voidable paste within the bladder may beinjected via the ureter under pressure. Optionally, a slowlybiodegradable, viscous or sticky pharmaceutical formulations of thepresent invention can be useful for its ability to adhere to an interiorwall of the bladder as needed so the drug is not flushed away when thebladder is emptied during urination.

A patient with OAB may need a temporary or chronic treatment for OAB anda direct administration of a medication comprising a compound of thepresent invention, the compound may be for example blebbistatin, with orwithout an “other drug” is an embodiment of the present invention andthis embodiment includes a direct instillation or deposition into theintravesical space of the bladder in the form of an immediate releasewith or without a portion of the formulation performing a slow-releaseof the medication.

Optionally, for the present invention, the intra-bladder chambermedication delivery can occur via a small pumping device drawingmedication from a drug reservoir containing a formulation capable ofbeing pumped at a controlled rate from the reservoir into the bladderchamber wherein pump device and reservoir of drug is located already inthe bladder chamber or outside the bladder (Fraser, M. O., Lavelle, J.P., Sacks, M. S., Chancellor, M. B. “The future of BladderControl-Intravesical Drug Delivery, a Pinch of Peper, and Gene Theraphy”Rev. Urol. 2002; 4(1), pp 1-11).

The medication to treat OAB comprises a compound of the presentinvention, the compound may be for example blebbistatin, optionally withan “other drug” in a suitable solution, semisolid, powder, foam, gel,slowly forming gel, powder, slow-release formulation of the medication,optionally wherein the medication comprises a pharmaceuticallyacceptable salt, ester, or prodrug, complex, suspension, or suppository.Optionally the formulation is a form or size or condition that preventsits elimination during urination or micturation.

Examples of routes of administration of a medication of the presentinvention include the following route and any other well-known in theart, for example: (1) an oral route as a tablet, capsule, solution,paste, or spray; (2) a parenteral route as an intravenous injection, asubcutaneous injection, an intramuscular injection, or a muscle depot;(3) a surgically installed inside the body anywhere may be a storagedevice pump which selectively delivers a pharmaceutical compositionsolution of a compound of the present invention by an osmotic means, bya compression means, a cannula means, or by diffusion means; (4) a nasalinhalation route as a spray; a transdermal patch route, a rectalsuppository as a semisolid, solid or liquid solution; (5) asubcutaneous, parenteral muscle depot or directly introduced into thebladder via a cannula or injection or any other means of directintroduction of said drug into the bladder.

Preferably a pharmaceutical composition of the present invention isadministered into the bladder lumen or intervesicular as a solution orintroduced therein as a formulation capable of functioning as aslow-drug release formulation. Optionally to minimize urinary losses ofthe entire drug dose consequent to urination, the formulation of anykind of rate of release, the formulation can be designed to adhere inpart to a portion of the bladder wall or lumen, the formulationcontaining biologically-compatible adhesive molecules known in the artof pharmaceutical excipients with adhesive qualities.

The scope of the present invention includes methods of administering amedication as a therapeutic pharmaceutical means for assisting in therelaxation of a patient's bladder smooth muscle for treating OAB(over-active bladder). Smooth muscle relaxants known in the art may beused as the “other drug”. This invention also relates to pharmaceuticalcompositions comprising said compounds and optionally another drug (“theother drug”) or a plurality of other drugs (“the other drugs”) andmethods of treating a lower urinary tract dysfunction using the same.The present invention is a method of treating a lower urinary tractdysfunction using a Myosin ATPase inhibitor compound and optionally,with an “other drug”(s), or an enantiomer, a pro-drug, a tautomer, apolymorph or a pharmaceutically acceptable salt thereof, said method oftreatment advantageously having a reduced side effect or an improvedtherapeutic effectiveness, or an improved patient compliance in takingtheir medication for the treatment of their lower urinary tracttreatment dysfunction(s).

The present invention includes methods of administering an “other drug”as a means for providing a therapeutic pharmaceutical for treating apatient with OAB. The other drug when combined with blebbistatin oranother Myosin II ATPase inhibitor may have additive, and preferablysynergistic effects. By treating an overactive bladder with twodifferent drug therapies, such as (a) a Myosin II ATPase inhibitor and(b) an “other drug”, the dose of each can be lower, advantageouslyreducing potential side-effects and reducing a development of metabolictolerance and a functional tolerance to the medications.

The “other drug” may be a drug known or that will be discovered tomodulate smooth muscle contractility, tonus or phasic tension, orrelaxation process biochemical pathways. Such modulation may arise whenthe “other drug” affects levels of cellular cyclic AMP or cyclic GMP,alters levels of synthesis or release of nitric oxide, carbon monoxide,hydrogen sulfide, oxygen, or carbon dioxide in and about bladder smoothmuscle cells and tissue. The “other drug” may reduce calcium ion influxinto smooth muscle cells, reducing calcium release or storage fromsmooth muscle intracellular organelles which may play a role in causingcalcium ion release or storage by intracellular organelle includingendoplasmic reticulum, sarcoplasmic reticulum, mitochondria, calciumbinding proteins such a calmodulin or the cell membrane (“glycocalax”)or other cellular membranes. The “other drug” may affect Na/Ca exchange;may block calcium channels; may increase or reduce sodium ionefflux/influx; may modulate the IP3 pathway; modulate prostaglandinsynthesis; modulate G proteins. The “other drug” may be a cholinergicantagonist such as a muscarinic or nictotinic anatagonist or induceincreased levels of an acetylcholinesterase enzyme or an adrenergicantagonist, an inducer of levels of a catechol-o-methyl transferase or amonoamine oxidase or blocker of acetylcholine synthesis or of neuronalrelease of acetylcholine near smooth muscle cells. The “other drug” mayreduce the duration or magnitude of smooth muscle cell membranedepolarization to effect smooth muscle relaxation.

Racemic or (−)-blebbistatin or another Myosin II ATPase inhibitor of thepresent invention when administered at the same time or having an effectin the same time period as an “other drug”, regardless of whether sameor separate pharmaceutical formulations, may surprisingly andadvantageously reduce the dosage by each drug needed for inducing smoothmuscle relaxation as well as prolonging the therapeutic duration ofsmooth muscle relaxation while beneficially reducing side effects thateither drug alone administered may cause. For the present inventionmethod of treating an overactive bladder of a patient, the method mayinclude administering a bladder smooth muscle relaxing agent.

A preferred compound of formula I for the present invention is racemicor (−)-blebbistatin or a Formula 1 or Formula II compound of the presentinvention. Said compound may be formulated alone or in combination withan “other drug” in a pharmaceutical formulation. The “other drug” may bea phosphodiesterase enzyme inhibitor (PDEI). During formation of cyclicAMP or cyclic GMP, a PDEI prevents breakdown of the cyclic monophosphatenucleotide thereby the smooth muscle cytoplasmic cyclic nucleotidelevels (i.e., cyclic AMP or cyclic GMP levels) rise higher and lastlonger, facilitating and prolonging smooth muscle relaxation. The PDEIsinclude cyclic GMP PDEI and cyclic AMP PDEI inhibitors. More than 20phosphodiesterase enzymes (PDE) are known and named PDE1 to PDE20. Thedistribution of PDEs differs in different cells and organs. The presentinvention's “other drug” may be a selective inhibitor of one or more ofthe following phosphodiesterase enzymes: PDE1, PDE2, PDE3, PDE4, PDE5,PDE6, PDE7, PDE8, PDE9, PDE10, PDE11, or PDE12. More preferably the“other drug” is a selective inhibitor of PDE1, PDE4 or PDE5. Mostpreferably the “other drug” is a selective inhibitor of PDE5. Preferred“other drug:” selective PDE5 inhibitors include sildenafil citrate,vardenafil, or tadalifil which slow hydrolysis of cyclic GMP formed bythe nitric oxide signaling pathway. Most preferably, when the “otherdrug” is a PFE5 inhibitor, then the “other drug” is vardenafil. Examplesrelatively nonselective PDE inhibitors include caffeine, theophylline,milrinone, and pentoxifylline which inhibit more than one type of PDEand thereby stabilize elevations in cyclic GMP or cyclic AMP in smoothmuscle cells depending upon whether CAMP or cGMP is being formed.

The term IC₅₀ applied to PDE inhibitor study means the concentration ofa PDE inhibitor (PDEI) that causes a 50% inhibition of the PDE'sactivity by whatever assay means that the IC50 is determined. Forexample, PDE5 selective inhibitor has a lower IC₅₀ for its selectedtarget, PDE5, than for its non-selected target PDE4. The selectivity ofa PDE inhibitor is preferably greater than about 10, more preferablygreater than about 100, and most preferably greater than about 1000. Apotent PDE inhibitor typically will have an IC₅₀ less than about 10micromolar, preferably less than about 1 micromolar, more preferablyless than 0.1 micromolar, and most preferably less than 0.01 micromolar.

For the present invention, an administration of a compound of theinvention, the compound may be for example, blebbistatin, from theinside chamber of the bladder is an effective means for relaxing abladder smooth muscle of a patient in order to treat OAB. As shown inFIG. 7B, blebbistatin can reduce both the baseline tone and peak phasiccontraction. Preferably the bladder's resting or peak phasic contractionare reduced by the treatment with a compound of the invention, thecompound may be for example, blebbistatin, and optionally along with the“other drug” by between about 5 to about 100 percent, to between about15 to about 100 percent, to between about 25 to 100 percent, preferablyto between about 25 to about 90 percent, more preferably to betweenabout 40 to about 80 percent, most preferably to between about 50 toabout 75 percent.

It is unexpected that a drug of the present invention reduces spasmodicbladder activity as evident in FIG. 3, FIG. 7A and FIG. 7B. When thebladder is in a contractile state induced by membrane depolarizationusing 60 mM potassium chloride (KCl) as shown in FIG. 1, or istransiently contracted by electric field stimulation (EFS) atfrequencies of 2 Hz to 16 Hz as in FIG. 5A, blebbistatin is an effectiveand useful means for inhibiting the contractility of the bladder.Similarly when a bladder is contracted by carbacol, the bladder smoothmuscle can still be relaxed by blebbistatin.

The efficacy of the present invention to relax a contracted bladder issurprising in view of the fact that an adult mouse bladder does notrelax when treated with blebbistatin whereas the rat bladder does relaxwhen exposed to blebbistatin.

IC₅₀ of a Compound of the Invention for Inhibition of Contraction of aRat Bladder Strip:

The in vitro IC₅₀ determination for a compound of the invention is basedon a test measuring the inhibition of a contraction by an adult ratisolated bladder muscle strip. Said test uses of rat bladder smoothmuscle strips is performed in the dark in a tissue chamber apparatus,optionally illuminated with a low intensity red lamp, since somecompounds of the invention are known to be light-sensitive to blue topurple or shorter wavelengths of light radiation.

Preferably, eight of the bladder muscle strips are employed in vitro ina first type of inhibition testing. Preferably, eight other bladdermuscle strips are employed in a second type of inhibition testing. Themuscles strips for each of the test types are obtained, one strip eachfrom eight animals, the animals being Sprague-Dawley rats of 300-375gram weight and purchased from Charles River Laboratories in the UnitedStates. A mean IC₅₀ determination per test compound per test type isbased on experimental data from 8 muscle test strips In the first typeof inhibition testing, the muscle strip in vitro bathed in aphysiological saline for an hour as defined in the present detaileddescription and then contracted with 1 micromolar (uM) carbacol(carbamylcholine), in the saline in a dark tissue chamber, which peaktension takes about 10 minutes. Representative 1 uM carbacol contractiondata are in FIG. 2. Several concentrations of the test compound areadded in increasing concentrations. For example, the present inventiontested 1 uM, 5 uM, and 10 uM racemic blebbistatin. The magnitude of amuscle strip contraction is measured when the magnitude is appears tohave reached a stable point which may take 10-40 minutes of testcompound exposure. One of ordinary skill in the art knows a mean IC₅₀can be calculated from such type one test data.

In the second test type, the muscle strip contractions can triggeredusing about 4 to 6 minutes of electric field stimulation (EFS) atfrequencies of 2, 4, 8, 16, and 32 as shown in FIG. 3. Then the EFScontractions are repeated in the presence of increasing concentrationsof the test compound. A test result is shown in FIG. 3 for testing with10 uM racemic blebbistatin. One of ordinary skill in the art knows anIC₅₀ can be calculated from such type two test data. The overall IC₅₀ ofa test compound defined as a calculated value midway between the typeone test mean measured IC₅₀ and the type two test mean measured IC₅₀.Said defined overall IC₅₀ value is a test compound's functional ratbladder contractility IC₅₀ for the present invention. It is intended forthe present invention that for each test compound, its overallfunctional rat bladder contractility IC₅₀ being defined can besystematically useful in a number of ways, including (a) to compare thepotency of two test compounds, (b) to discovery a useful compound of thepresent invention and (c) to conduct infringement or anticipation by acompound. Further details of such IC₅₀ determinations are taught in thedetailed description.

Detailed methods for studying in vitro changes in the contractility ofrat bladder smooth muscle caused by a Myosin II ATPase inhibitor, suchas blebbistatin, may be found in Zhang, X, Kuppam, D S R, Melman, A,DiSanto, M E. “In vitro and In vivo relaxation of urinary bladder smoothmuscle by the selective Myosin II inhibitor, blebbistatin” BJU Internat.e-publication 2010, journal publication, 2011; Volume 107, Issue 2,pages 310-317.

Example 1 In Vitro Test Response of Rat Bladder and Human Bladder SmoothMuscle Strip to Blebbistatin

17 male Sprague Dawley rats along with multiple human bladder smoothmuscle strips obtained from an open prostatectomy were used for racemicblebbistatin organ bath studies. Awake cystometry was performed on aseparate set of 5 rats in both the presence and absence ofintravesically delivered racemic blebbistatin. The effect of racemicblebbistatin on pharmacologically and nerve-mediated bladder SMcontraction was determined. The influence of racemic blebbistatin onurodynamic properties was also assessed. Results: Racemic blebbistatincompletely relaxed both KCl and carbachol induced rat detrusor andendothelin-1 induced human bladder (limited to base) contraction in adose-dependent manner. Pre-incubation with 10 μM racemic blebbistatinattenuated carbachol responsiveness by 65% while blocking electricalfield stimulation-induced bladder contraction reaching 50% inhibition at32 Hz. Basal tone and amplitude of spontaneous contraction were alsosignificantly blunted. Urodynamic parameters, consistent with OAB, wereobviously ameliorated by racemic blebbistatin intravesical infusion. Thenovel data of the present invention establish that racemic blebbistatinpotently strongly relaxes both rat and human bladder contraction inducedby various physiological agonists. In vivo tests showed that nanomolardoses of racemic blebbistatin significantly alter urodynamic parametersto a less active bladder.

Experimental materials and methods: All chemical were from Sigma (St.Louis, USA) except (±) blebbistatin (racemic blebbistatin) was fromTocris (Ellisville, Mo.). A stock solution of racemic blebbistatin wasmade in DMSO (dimethylsulphoxide; the other substances were dissolveddaily in double distilled water. Control experiments showed that thefinal concentrations (1/1000-3/1000) of DMSO used in these studies didnot significantly modify the relaxation response induced by racemicblebbistatin. Due to the known light sensitivity of blebbistatin, it wasalways kept in the dark in the refrigerator until just prior to usage.During the experiment, the organ bath chambers were covered. Humanbladder (base) samples were obtained from simple open benignprostatectomy surgery with informed consent and approval of theInstitutional Review Boards of Montefiore Medical Center and the AlbertEinstein College of Medicine. Rat bladder body and aorta were obtainedfrom 17 (275-300 gram) adult male Sprague-Dawley (SD) rats (CharlesRiver; Raleigh, N.C.). All animal studies were approved by the AnimalInstitute Committee of the Albert Einstein College of Medicine.

It was determined that the active (−) enantiomer form of blebbistatinwas equipotent to the racemic blebbistatin mixture in the in vitrostudies and that the inactive (+) enantiomer form of blebbistatin formdid not induce significant bladder relaxation.

In vitro organ bath studies: The in vitro contractility studies wereperformed as previously described by Zhang X, Kuppam D, Aydin M, MelmanA, DiSanto M. “In vitro and in vivo relaxation of corpus cavernosumsmooth muscle by the selective myosin II inhibitor, blebbistatin.” J SexMed 2009; 6:2661-71, Sandhu K S, Chua R G, Zhang X, et al. “Regionalheterogeneity in expression of the sphingosine-1-phosphate pathway inthe female rat lower urinary tract”; Am J Obstet Gynecol 2009;200:576-7; Chua R G, Calenda G, Zhang X, et al. “Testosterone regulateserectile function and Vcsa1 expression in the corpora of rats”; Mol CellEndocrinol 2009; 303:67-73. Bladder strips were mounted longitudinallyand aortic rings horizontally in a 5 ml organ bath—Multi-Myograph Model810MS (bladder) or Model 610M (aorta) (Danish Myo Technology; Aarhus,Denmark). The myograph was connected in line to a PowerLab 4/30 DataAcquisition System (ADInstruments; Colorado Springs, Colo.) and in turnto a Dual-Core processor Pentium computer for real-time monitoring ofphysiological force. The smooth muscle strips were equilibrated at least1 hour in Krebs-Henseleit (Krebs) buffer at 37° C. with continuousbubbling of 95% O2 and 5% CO2 and buffer was changed every 15 minutes(min). Strips were continuously adjusted to resting tension (0.5 g forrat bladder and 1 g for human bladder (Yamamoto M, Harm S C, Grasser WA, Thiede M A. “Parathyroid hormone-related protein in the rat urinarybladder: a smooth muscle relaxant produced locally in response tomechanical stretch.” Proc Natl Acad Sci USA 1992; 89:5326-30. Afterequilibration, rat detrusor was contracted with 60 mM KCl and the forceinduced by cumulative concentrations (10-5-10-1 M) of carbachol (CC) orelectrical field stimulation (EFS) at varying frequencies of 2-32 Hz,pulse duration 1.5 ms, train 5 s, and 80 V, was normalized to that KClresponse. Next, all strips were pre-contracted with 60 mM KCl, or 1 μMCC (rat bladder), or 20 nM endothelin-1 (ET-1) (human bladder) andallowed to reach stable tension and then the relaxant effects ofincreasing doses of blebbistatin were evaluated. For rat detrusor, afterpre-incubation with blebbistatin (10 μM) for 30 min, its inhibitoryeffect on CC (1 μM) induced contractility or aforementioned EFS mediatedcontractility was also tested. Additionally, the influence ofblebbistatin (10 μM) on rat detrusor spontaneous activity and basal tonewas evaluated.

Example 2 In Vivo Urodynamic Studies in Rats

Cystometric evaluation of bladder function was performed as previouslydescribed on a separate set of 5 rats. See Malmgren A, Andersson P O,Uvelius B. “Bladder function in rats with short- and long-term diabetes;effects of age and muscarinic blockade” J Urol 1989; 142:1608-14; MelmanA, Zotova E, Kim M, et al. “Longitudinal studies of time-dependentchanges in both bladder and erectile function afterstreptozotocin-induced diabetes in Fischer 344 male rats” BJU Int 2009,104:1292-300; Suadicani S O, Urban-Maldonado M, Tar M T, Melman A, SprayD C. “Effects of ageing and streptozotocin-induced diabetes onconnexin43 and P2 purinoceptor expression in the rat corpora cavernosaand urinary bladder” BJU Int 2009; 103:1686-93. Under anesthesia, a PE50 catheter with a cuff was surgically inserted into the bladder domeand exited through an orifice made in the back of the animal.Cystometrical analyses were performed two days after the surgery, as weshowed this to be an optimal period for recovery and investigation. Thebladder catheter was connected to a 2-way valve that is, in turnconnected to a pressure transducer as well as an infusion pump (modelPHD 2000, Harvard Instruments; MA, USA). Rat bladder was infused withvehicle for 30 min and then cystometry was performed. After that, thesame rat was treated with 250 nmols blebbistatin for the same period andcystometry was similarly repeated. One rat had bladder over-activitypossibly due to post-surgical infection and thus blebbistatin was keptin the bladder for only about 5 min. The rate of infusion of roomtemperature saline was set at 10 ml/h. Bladder activity was continuouslyrecorded after the first micturition, and subsequently at least 2 h ofdata were recorded from each rat. Statistical analysis: Results areexpressed as mean±SEM for n experiments. Statistical analysis wasperformed using either the Student's t-test (when two sample treatmentswere being compared) or using ANOVA when multiple means were compared.p<0.05 was considered significant.

Results: In rat bladder smooth muscle strips in vitro mounted in atissue bath, both KCl and carbacol (CC) produced phasic-type tensionincreases. CC dose-response curves were normalized to KCl elicited forceand averaged. About 50% of maximum contraction (around 125% of KClresponse) was reached at 1 μm and this submaximal contraction was chosenfor later experiments.

FIG. 1, FIG. 2, and FIG. 3. tracings of bladder strip tension versustime are representative force tracings and rat detrusor (the “as awhole, the bladder muscle”) when pre-contracted with 60 mM KCl or 1 μmagonist (CC) can be dose-dependently and completely relaxed by BLEB. Onaverage, blebbistatin relaxed both KCl (FIG. 1) and CC (FIG. 2, FIG. 3)evoked detrusor contraction by 20-30% at 1 μm, 80-100% at 5 μm and morethan 100% at 10 μm.

When the rat detrusor muscle (smooth muscle bladder strip) waspre-incubated with 10 μm blebbistatin for 30 min, it not only stronglyblunted both CC (FIG. 4A and FIG. 4B) and EFS (FIG. 5A and FIG. 5B)induced detrusor contraction amplitude, it also significantly loweredthe basal tone by around 300 mg. In fact, it attenuated CCresponsiveness by about 65% (FIG. 4A and FIG. 4B), while blockingEFS-induced stimulation at all frequencies (FIG. 5A and FIG. 5B), butwas more pronounced at higher frequencies reaching 50% inhibition at 32Hz.

The potent inhibitory profile of blebbistatin was further confirmed inhuman detrusor smooth muscle obtained from a BPH/LUTS patient bladderbase near the bladder neck, as illustrated in FIG. 6A and FIG. 6B. A 1micromolar (μM), blebbistatin produced 60% relaxation of human bladderthat is being pre-contracted with endothelin-1 (ET-1), one of thestrongest vasoconstrictors known (20 nM ET-1) and nearly 100% relaxationat 3 μm.

Animal detrusor smooth muscle strips develop spontaneous activity Asshown in FIG. 7A and FIG. 7B and Table 1, blebbistatin (10 μm) causesabout a 60% decrease ion the amplitude of spontaneous smooth musclecontractions in rat bladder muscle strips (from 287.08±33.23 milligrams(mg) to 118±19.33 mg.) as well as a significant relaxation to the tonus(baseline tension). See FIGS. 4A and 5A. (FIGS. 3A & C).

TABLE 1 Effect of blebbistatin on spontaneous contractions of ratbladder strips Frequency Treatment Basal Tone (mg) Amplitude (mg)(cycles/min) Without 590 +/− 23 ** 290 +/− 33 4.6 +/− 1.1 blebbistatinWith 310 +/− 16 120 +/− 19 5.5 +/− 1.2 blebbistatin n = 15 strips from 6different rats, ** <0.01 vs with blebbistatin.

Example 3 Experimental In Vivo Instillation of 250 NanomolarBlebbistatin into the Normal Awake Rat Bladder

FIG. 8A shows typical rat cystometric tracings in which the bladder hasbeen pre-treated with vehicle (left panels) or blebbistatin (rightpanels) for 30 min. The urodynamic parameters are summarized in Table 2below. Table 2 shows that BLEB significantly increased bladder capacity,micturition volume and compliance while micturition frequency wasobviously decreased. Intravesical pressure was not significantlyinfluenced by blebbistatin. FIG. *B is the uroflow tracing of the ratwith overactive bladder treated with vehicle (upper panel) orblebbistatin (lower panel). Consistent with human BPH/LUTS bladderstrips (obtained from simple human prostatectomy operations) in vitroobservations, rats with OAB are more sensitive to blebbistatin withvoiding frequency declining by over 50%.

TABLE 2 Impact of blebbistatin on UroDynamic Parameters of Awake Rats (n= 5) Treatment BC (ml) MV (ml) RV (ml) Vehicle 0.78 +/− 0.15 0.76 +/−0.15 0.02 +/− 0.01 Blebbistatin  1.4 +/− 0.16 *  1.4 +/− 10 * 0.03 +/−0.01 Treatment BP (cm H20) MP (cmH20) Bcom (ml) MF (ml) Vehicle 7.1 +\−0.91 40 +/− 12 0.16 +/− 0.01  13 +/− 3.3 Blebbistatin 6.9 +/− 1.8 42 +/−13 0.28 +/− 0.02 7.2 +/− 1.3 *

Bladder function was evaluated using the following urodynamic criteria:bladder capacity: (BC=volume of infused saline discharged atmicturation); micturation volume (MV=the volume of urine dischargedduring micturation); residual volume (RV=volume of infused saline minusmicturation volume); basal pressure (BP=the lowest average bladderpressure recorded during cystometry); micturation pressure (MP=peakbladder pressure during micturation); Bcom=bladder compliance;MF=micturation frequency; *<0.05 vs vehicle.

Studies reported regarding the present invention indicate that in vitroin a tissue bath, the smooth muscle rat bladder strips generated fastphasic-type contraction elicited by potassium chloride (KCl)depolarization or carbacol (CC) evoked activation of the muscarinicsystem (FIG. 1, FIG. 2 and FIG. 3). Neither method of inducing a phasiccontraction in the smooth muscle bladder strip influenced the means bywhich the smooth muscle bladder strip relaxed.

In vitro relaxation in response to blebbistatin was dose-dependent: withbasal tension decreasing by 20-30% at 1 μm, 80-100% at 5 μm and over100% at 10 lam. In vitro pre-incubation of bladder strips with BLEB (10μM) for 30 minutes attenuated a carbacol-mediated bladder stripcontraction by about 65% while EFS-evoked contraction was attenuatedfrequency by frequency reaching at 32 Hz by about 50%. Peak contractionforce and steady-state tension were strongly reduced (FIG. 4A and FIG.4B). The time needed for blebbistatin to relax the muscle strip in vitroin a tissue bath was 30-60 minutes whereas muscarinic receptorantagonists such as atropine inhibit relaxation in 3-5 minutes.

In in vitro experiments related to the present invention, rat smoothmuscle bladder strips exhibited spontaneous contractions which 10 uMBLEB significantly reduced in spontaneous contraction amplitude and inbasal tone (FIG. 7A and FIG. 7B and Table. Thus, the present experimentsare support the utility of the present invention as therapeutic meansfor treating OAB.

Human bladder base smooth muscle tissue from open prostatectomyoperations was studied to test the utility of the present invention.BLEB induced a strong, dose-dependent relaxation of human bladder invitro tissue (FIG. 6A and FIG. 6B). In vitro, blebbistatin at a dose of1 μM caused a 60% relaxation of human bladder tissue that had beenpre-contracted with Endothelian-1 (ET-1) a strong vasoconstrictor. ABPH/LUTS bladder may have an enhanced response to blebbistatin due todistension or inflammation of the overactive bladder (Lavelle J P,Meyers S A, Ruiz W G, Buffington C A, Zeidel M L, Apodaca G. “Urothelialpathophysiological changes in feline interstitial cystitis: a humanmodel” Am J Physiol Renal Physiol 2000; 278: F540-F553).

In one experiment testing the utility of the present invention, 250nanomolar BLEB was instilled into the bladder of a normal awake rat. Theintra-bladder administration of blebbistatin effectively inhibitedcontractility, increased bladder capacity, volume of micturition andbladder compliance while decreasing the frequency of urination (FIG. 8A,FIG. 8B and Table. 2). In vivo, in an experiment with a rat sufferingOAB, BLEB was more effective than in the rat with a normal functioningbladder.

EQUIVALENTS

Numerous modifications and alternative embodiments of the presentinvention will be apparent to those skilled in the art in view of theforegoing description. Accordingly, this description is to be construedas illustrative only and is for the purpose of teaching those skilled inthe art the best mode for carrying out the present invention. Details ofthe structure may vary substantially without departing from the spiritof the invention, and exclusive use of all modifications that comewithin the scope of the appended claims is reserved. It is intended thatthe present invention be limited only to the extent required by theappended claims and the applicable rules of law.

All literature and similar material cited in this application,including, patents, patent applications, articles, books, treatises,dissertations and web pages, regardless of the format of such literatureand similar materials, are expressly incorporated by reference in theirentirety. In the event that one or more of the incorporated literatureand similar materials differs from or contradicts this application,including defined terms, term usage, described techniques, or the like,this application controls. The section headings used herein are fororganizational purposes only and are not to be construed as limiting thesubject matter described in any way.

While the present inventions have been described in conjunction withvarious embodiments and examples, it is not intended that the presentteachings be limited to such embodiments or examples. On the contrary,the present inventions encompass various alternatives, modifications,and equivalents, as will be appreciated by those of skill in the art.The claims should not be read as limited to the described order orelements unless stated to that effect. It should be understood thatvarious changes in form and detail may be made without departing fromthe scope of the appended claims. Therefore, all embodiments that comewithin the scope and spirit of the following claims and equivalentsthereto are claimed.

1. A method of treating an overactive bladder in a patient, which methodcomprises: administering to said patient in an active form an effectiveamount of a Myosin II ATPase inhibitor compound of Formula (I) or apharmaceutically-acceptable salt, a racemic mixture, an enantiomer, or aprodrug thereof, to treat the overactive bladder in the patient,

including: selecting an X from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, carboxy, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, carbamyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, methylamino, amino, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, acetamido,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,acetonyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, phenyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, napthyl,benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl,alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparaginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, the X optionallysubstituted at a carbon atom with a substituent W selected from thegroup consisting of methyl, hydroxy, fluoro, chloro, bromo, iodo, nitro,cyano, thio, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, phenyl, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, carbamyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkoxy, amino, methylamino,napthyl, aminomethyl, amino(C₁₋₆)alkyl, benzyl, acetamido,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,acetoxy, acetonyl, carb(C₁₋₆)alkoxy, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, cyclopropyl, (C₃₋₆)cycloalkyl, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₄₋₇)cycloalkenyl,C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,(C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl,asparaginyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andthe X optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent Q selected from the groupconsisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic, (C₄₋₁₀)aryl,(C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; and selecting a Y from the groupconsisting of hydrogen, methyl, hydroxy, fluoro, chloro, bromo, iodo,nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, carbamyl, chloromethyl, fluoromethyl,trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, aminomethyl,methylamino, amino, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, phenyl, benzyl, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, napthyl, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, the Y optionallysubstituted at a carbon atom with a substituent Z selected from thegroup consisting of methyl, hydroxy, fluoro, chloro, bromo, iodo, nitro,cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, carbamyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, chloromethyl, fluoromethyl,trifluoromethyl, thio(C₁₋₆)alkyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, acetonyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl,asparaginyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andthe Y optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent T selected from the groupconsisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl, phenyl,C₂₋₆)alkynl, (C₁₋₆)haloalkyl, benzyl, napthyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,and (C₄₋₈)heteroaryl.
 2. The method according to claim 1, includingselecting said Myosin II ATPase inhibitor compound of Formula I inaddition having an IC₅₀ of between about 0.0001 to about 50 micromolarbased upon an in vitro determination.
 3. (canceled)
 4. The methodaccording to claim 1, including: selecting the X from the groupconsisting of hydrogen, amino, methyl, hydroxy, fluoro, chloro, bromo,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino(C₁₋₆)alkyl, acetonyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,carb(C₁₋₆)alkoxy, (C₁₋₆)alkylaminoketo, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,acetoxy, acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, and acetonyl, the X optionally substituted at a carbonatom with the substituent W selected from the group consisting ofmethyl, hydroxy, fluoro, chloro, acetyl, (C₁₋₆)alkyl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy; and selecting the Y from thegroup consisting of hydrogen, amino, methyl, hydroxy, fluoro, chloro,bromo, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino(C₁₋₆)alkyl, acetonyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,carb(C₁₋₆)alkoxy, (C₁₋₆)alkylaminoketo, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,acetoxy, acetamido, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, and acetonyl, the Y optionally substituted at a carbonatom with the substituent Z selected from the group consisting ofmethyl, hydroxy, fluoro, chloro, acetyl, (C₁₋₆)alkyl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy.
 5. The method, according to claim4, including: selecting the X from the group consisting of hydrogen,methyl, fluoro, chloro, bromo, acetyl, (C₁₋₆)alkyl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, hydroxy, amino, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl, and (C₁₋₆)haloalkoxy; andselecting the Y from the group consisting of hydrogen, methyl, fluoro,chloro, bromo, acetyl, (C₁₋₆)alkyl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, hydroxy, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, amino,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, and (C₁₋₆)haloalkoxy.
 6. (canceled)
 7. Themethod according to claim 1, including: selecting the X from the groupconsisting of hydrogen, methyl, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, napthyl, benzyl, and (C₄₋₈)heteroaryl, the Xoptionally substituted at a carbon atom with a the substituent Wselected from the group consisting of methyl, hydroxy, fluoro, chloro,acetyl, (C₁₋₆)alkyl, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino(C₁₋₆)alkyl, amino,(C₁₋₆)alkylamino, and (C₁₋₆)haloalkoxy, and the X optionally substitutedat a primary nitrogen atom or at a secondary nitrogen atom with thesubstituent Q selected from the group consisting of methyl, (C₁₋₆)alkyl,cyclopropyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, phenyl,benzyl, napthyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,(C₄₋₈)heterocyclic, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl;and selecting the Y from the group consisting of hydrogen, methyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, and(C₄₋₈)heteroaryl, the Y optionally substituted at a carbon atom with thesubstituent Z selected from the group consisting of methyl, fluoro,chloro, hydroxy, acetyl, (C₁₋₆)alkyl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,amino, and (C₁₋₆)haloalkoxy, the Y optionally substituted at a primarynitrogen atom or at a secondary nitrogen atom with the substituent Tselected from the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic,(C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl.
 8. (canceled) 9.(canceled)
 10. The method according to claim 1, including selecting theMyosin II ATPase inhibitor compound of Formula (1) from the groupconsisting of racemic blebbistatin, (−)-blebbistatin or apharmaceutically-acceptable salt, or a prodrug thereof.
 11. A method oftreating an overactive bladder in a patient, which method comprises:administering to said patient in an active form an effective amount of aMyosin II ATPase inhibitor compound of formula (II), or apharmaceutically-acceptable salt, a racemic mixture, an enantiomer, or aprodrug thereof, to treat the overactive bladder in the patient,

including: selecting an X from the group consisting of methyl, nitro,cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, amino, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl,trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl,methylamino, aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, acetoxy, (C₁₋₆)alkylaminoketo,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆) alkyl-O—C(═O(C₁₋₆)hydroxyalkyl, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, cyclopropyl,(C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl,benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl,alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, the X optionallysubstituted at a carbon atom with a substituent W selected from thegroup consisting of methyl, hydroxyl, fluoro, chloro, bromo, iodo,nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, thio, (C₁₋₆)alkoxy, methylthio, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, acetoxy,trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino,methylamino, aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl, acetamido,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, carb(C₁₋₆)alkoxy,(C₁₋₆)alkoxy(C₁₋₆)alkyl, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, acetonyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl,asparginyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, andthe X optionally substituted at a primary nitrogen atom or at asecondary nitrogen atom with a substituent Q selected from the groupconsisting of methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, benzyl, napthyl, (C₄₋₈)heterocyclic, (C₄₋₁₀)aryl,(C₄₋₁₀)haloaryl, and (C₄₋₈) heteroaryl; selecting a Y from the groupconsisting of methyl, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, amino, glycinyl,alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, the Y optionallysubstituted at a carbon atom with a substituent Z selected from thegroup consisting of methyl, hydroxyl, fluoro, chloro, bromo, iodo,nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, methylthio, thio, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl, carbamyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, amino, methylamino, aminomethyl,amino(C₁₋₆)alkyl, acetoxy, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl acetamido, cyclopropyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, carb(C₁₋₆)alkoxy,napthyl, phenyl, benzyl, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, benzyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, acetonyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, ((C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,(C₄₋₈)heteroaryl, C₄₋₁₀)aryl, glycinyl, alanyl, serinyl, valinyl,leucinyl, isolecinyl, prolinyl, methionyl, phenylalanyl, tyrosinyl,tryptophanyl, threonyl, cystinyl, asparginyl, lysinyl, histidinyl,arginyl, aspartyl, and glutamyl, and the Y optionally substituted at aprimary nitrogen atom or at a secondary nitrogen atom with a substituentT selected from the group consisting of methyl, (C₁₋₆)alkyl,cyclopropyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl,(C₃₋₇)cycloalkyl, phenyl, (C₄₋₇)cycloalkenyl, benzyl, (C₄₋₁₀)aryl,napthyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; andhaving an atom of the X in a chemical bond with an atom of the Y forminga ring ortho-fused to a ring of Formula (II).
 12. The method accordingto claim 11, including selecting said Myosin II ATPase inhibitorcompound of Formula II in addition, having an IC₅₀ of about 0.0001 toabout 50 micromolar, the IC₅₀ based upon an in vitro IC₅₀ determination.13. (canceled)
 14. (canceled)
 15. The method of claim 11, includingselecting the ortho-fused ring from the group consisting of thiophene,pyrrole, furan, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, andpyridazine, optionally substituted with a substituent W, the substituentW selected from the group consisting of methyl, hydroxyl, fluoro,chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy,and optionally substituted with a substituent Z, the substituent Zselected from the group consisting of methyl, hydroxyl, fluoro, chloro,bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy.16. A method of treating an overactive bladder in a patient, comprising:administering to said patient in an active form an effective amount of aMyosin II ATPase inhibitor compound, or a pharmaceutically-acceptablesalt, a racemic mixture, an enantiomer, or a prodrug thereof, to treatthe overactive bladder of the patient.
 17. A method according to claim16, including selecting the Myosin II ATPase inhibitor compound inaddition, in its active form, having an IC₅₀ of about 0.0001 to about 50micromolar, the IC₅₀ based upon an in vitro determination. 18.(canceled)
 19. The method according to claim 16, which furthercomprises: administering a drug selected from the group consisting of aPDE5 inhibitor drug and an antimuscarinic drug, to treat the overactivebladder of the patient.
 20. The method according to claim 19, furthercomprising: administering an effective amount of a Myosin II ATPaseinhibitor, optionally a PDE5 inhibitor drug, and optionally anantimuscarinic drug to the overactive bladder directly.
 21. A Myosin IIATPase inhibitor compound of Formula (I) or apharmaceutically-acceptable salt, a racemic mixture, an enantiomer, or aprodrug thereof, comprising:

wherein an X is selected from the group consisting of hydroxy, fluoro,chloro, bromo, iodo, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, methyl, nitro, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, carbamyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, methylamino, amino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, acetoxy,(C₁₋₆)alkylamino(C₁₋₆)alkyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetamido, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,cyclopropyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,acetonyl, phenyl, benzyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,cyclopropyl, napthyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,(C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl, valinyl, leucinyl,isolecinyl, prolinyl, methionyl, phenylalanyl, tyrosinyl, tryptophanyl,threonyl, cystinyl, asparaginyl, lysinyl, histidinyl, arginyl, aspartyl,and glutamyl, wherein the X is optionally substituted at a carbon atomwith a substituent W selected from the group consisting of methyl,hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy,acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, thio,(C₁₋₆)alkoxy, methylthio, thiomethyl, C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, carbamyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, amino, methylamino, acetonyl, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₄₋₁₀)aryl,acetoxy, phenyl, napthyl, (C₁₋₆)alkylamino(C₁₋₆)alkyl, benzyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁-6)alkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, acetamido,(C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl, valinyl, leucinyl,isolecinyl, prolinyl, methionyl, phenylalanyl, tyrosinyl, tryptophanyl,threonyl, cystinyl, asparaginyl, lysinyl, histidinyl, arginyl, aspartyl,and glutamyl; and wherein the X is optionally substituted at a primarynitrogen atom or at a secondary nitrogen atom with a substituent Qselected from the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, phenyl,benzyl, napthyl, (C₄₋₇)cycloalkenyl, (C₄₋₈)heterocyclic, (C₄₋₁₀)aryl,(C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; and wherein a Y is selected fromthe group consisting of hydroxy, fluoro, chloro, bromo, cyano, carboxy,carbethoxy, acetyl, (C₂₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,methythio, (C₁₋₆)alkoxy, thio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, iodo, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl, amino, methylamino,aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, acetamido, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, acetoxy, carb(C₁₋₆)alkoxy, cyclopropyl,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,(C₁₋₆)hydroxyalkyl, acetonyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, nitro,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl,napthyl, benzyl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl,glycinyl, alanyl, serinyl, valinyl, leucinyl, isolecinyl, prolinyl,methionyl, phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl,asparginyl, lysinyl, histidinyl, arginyl, aspartyl, and glutamyl,wherein the Y is optionally substituted at a carbon atom with asubstituent Z selected from the group consisting of methyl, hydroxy,fluoro, chloro, bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl,(C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,methylthio, thio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, carbamyl, fluoromethyl, amino, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, acetoxy, (C₁₋₆)alkylamino,carb(C₁₋₆)alkoxy, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, acetonyl,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, phenyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, napthyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,benzyl, (C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl, valinyl, leucinyl,isolecinyl, prolinyl, methionyl, phenylalanyl, tyrosinyl, tryptophanyl,threonyl, cystinyl, asparaginyl, lysinyl, histidinyl, arginyl, aspartyl,and glutamyl, and wherein the Y is optionally substituted at a primarynitrogen atom or at a secondary nitrogen atom with a substituent Tselected from the group consisting of methyl, (C₁₋₆)alkyl, cyclopropyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, phenyl,napthyl, (C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, and benzyl.
 22. The compound ofFormula (I) according to claim 21, wherein the X is selected from thegroup consisting of hydroxy, thio, fluoro, chloro, acetyl, (C₂₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, acetoxy, bromo,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, nitro, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, acetamido, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylaminoketo, carb(C₁₋₆)alkoxy, (C₁₋₆)hydroxyalkyl,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, and acetonyl, the X optionallysubstituted at a carbon atom with the substituent W selected from thegroup consisting of methyl, hydroxy, fluoro, chloro, acetyl,(C₁₋₆)alkyl, methoxy, thio, (C₁₋₆)alkoxy, methylthio, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl,trifluoromethyl, (C₁₋₆)haloalkyl, and (C₁₋₆)halo alkoxy; and wherein theY is selected from the group consisting of hydroxy, fluoro, chloro,acetyl, (C₂₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, bromo,chloromethyl, fluoromethyl, trifluoromethyl, nitro (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, carbamyl, amino, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, acetamido, (C₁₋₆)alkylaminoketo,acetoxy, carb(C₁₋₆)alkoxy, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkanoyl, acetonyl,and (C₁₋₆)alkylcarboxy, the Y optionally substituted at a carbon atomwith the substituent Z selected from the group consisting of methyl,hydroxy, fluoro, chloro, acetyl, (C₁₋₆)alkyl, methoxy, (C₁₋₆)alkoxy,thio, methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, and(C₁₋₆)haloalkoxy.
 23. The compound of Formula (I) according to claim 22,wherein the X is selected from the group consisting of ethyl, hydroxy,fluoro, chloro, acetyl, (C₂₋₆)alkyl, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, nitro, thio(C₁₋₆)alkyl,chloromethyl, fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl,(C₁₋₆)haloalkoxy, bromo, amino, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino; and wherein the Y is selected from the groupconsisting of hydroxy, fluoro, chloro, acetyl, (C₂₋₆)alkyl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, amino, amino(C₁₋₆)alkyl, nitro, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, and (C₁₋₆)haloalkoxy.
 24. The compound of Formula (I)according to claim 22, wherein the X is selected from the groupconsisting of ethyl, ethoxy, methylthio, ethylthio, propylthio,fluoromethyl, trifluoromethyl, and trifluromethoxy, amino(C₁₋₆)alkyl,amino, (C₁₋₆)alkylamino; and wherein the Y is selected from the groupconsisting of hydrogen, ethyl, ethoxy, methylthio, nitro, ethylthio,propylthio, fluoromethyl, trifluoromethyl, trifluromethoxy, amino,amino(C₁₋₆)alkyl, and (C₁₋₆)alkylamino.
 25. The compound of Formula (I)according to claim 21, wherein the X is selected from the groupconsisting of a methyl, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl, the Xoptionally substituted at a carbon atom with the substituent W selectedfrom the group consisting of hydrogen, hydroxy, fluoro, chloro, acetyl,(C₁₋₆)alkyl, methoxy, (C₁₋₆)alkoxy, thio, methylthio, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl,trifluoromethyl, (C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy, and the Xoptionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with the substituent Q selected from the group consistingof hydrogen, (C₁₋₆)alkyl, cyclopropyl, napthyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, phenyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, benzyl, (C₄₋₁₀)aryl, (C₄₋₈)heterocyclic,(C₄₋₁₀)haloaryl, and (C₄₋₈)heteroaryl; and wherein the Y is selectedfrom the group consisting of a cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl, the Yoptionally substituted at a carbon atom with the substituent Z selectedfrom the group consisting of hydrogen, methyl, hydroxy, fluoro, chloro,acetyl, (C₁₋₆)alkyl, nitro, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy,and optionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with the substituent T selected from the group consistingof hydrogen, methyl, (C₁₋₆)alkyl, cyclopropyl, benzyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl,phenyl, napthyl, (C₄₋₈)heterocyclic, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and(C₄₋₈)heteroaryl.
 26. A Myosin II ATPase inhibitor compound of Formula(II), or a pharmaceutically-acceptable salt, a racemic mixture, anenantiomer, or a prodrug thereof, comprising:

wherein an X is selected from the group consisting of methyl, cyano,carboxy, acetyl, carbethoxy, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl,methoxy, (C₁₋₆)alkoxy, thio, methylthio, amino, thiomethyl,(C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, acetoxy,trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl,methylamino, aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂ (C₁₋₆)alkylaminoketo, carb(C₁₋₆)alkoxy,(C₁₋₆)hydroxyalkyl, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl,(C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl, cyclopropyl,phenyl, benzyl, napthyl, (C₃₋₆)cycloalkyl, acetonyl, acetamido,(C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic,(C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl, valinyl, leucinyl,isolecinyl, prolinyl, methionyl, phenylalanyl, tyrosinyl, tryptophanyl,threonyl, cystinyl, asparginyl, lysinyl, histidinyl, arginyl, aspartyl,and glutamyl, wherein the X is optionally substituted at a carbon atomwith a substituent W selected from the group consisting of hydrogen,methyl, hydroxy, fluoro, chloro, bromo, iodo, nitro, cyano, amino,carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl,methoxy, (C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, carbamyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, methylamino, aminomethyl,amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino,(C₁₋₆)alkylamino(C₁₋₆)alkyl, phenyl, (C₁₋₆)alkylaminoketo, acetoxy,acetamido, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carb(C₁₋₆)alkoxy,(C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl, (C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl,(C₁₋₆)hydroxyalkyl, benzyl, (C₁₋₆)alkylcarboxy, acetonyl,(C₁₋₆)alkanoyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl, (C₁₋₆)alkoxy(C₁₋₆)alkyl,cyclopropyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, (C₄₋₁₀)aryl, napthyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenylalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, and wherein the Xis optionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with a substituent Q selected from the group consisting ofhydrogen, methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl,(C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, phenyl, (C₄₋₇)cycloalkenyl, benzyl,napthyl, (C₄₋₈)heterocyclic, (C₄₋₁₀)aryl, (C₄₋₁₀)haloaryl, and(C₄₋₈)heteroaryl; and wherein the Y is selected from the groupconsisting of hydrogen, hydroxy, methyl, fluoro, chloro, bromo, iodo,nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, methoxy, (C₁₋₆)alkoxy, thio, methylthio, thiomethyl,(C₁₋₆)alkylthio, amino, thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl,trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy, carbamyl,methylamino, aminomethyl, amino(C₁₋₆)alkyl, amino[(C₁₋₆)alkyl]₂,(C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,(C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, carbamyl, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, (C₁₋₆)alkylcarboxy,(C₁₋₆)alkanoyl, acetonyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, acetamido, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, benzyl,(C₄₋₁₀)haloaryl, (C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl,serinyl, valinyl, leucinyl, isolecinyl, prolinyl, methionyl,phenyllalanyl, tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl,lysinyl, histidinyl, arginyl, aspartyl, and glutamyl, wherein the Y isoptionally substituted at a carbon atom with the substituent Z selectedfrom the group consisting of hydrogen, methyl, hydroxy, fluoro, chloro,bromo, iodo, nitro, cyano, carboxy, carbethoxy, acetyl, (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy, methylthio, (C₁₋₆)alkoxy, thio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, trifluoromethyl, (C₁₋₆)haloalkyl, (C₁₋₆)haloalkoxy,carbamyl, amino, methylamino, aminomethyl, amino(C₁₋₆)alkyl,amino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylamino, (C₁₋₆)alkylamino(C₁₋₆)alkyl,carbamyl, (C₁₋₆)alkylamino[(C₁₋₆)alkyl]₂, (C₁₋₆)alkylaminoketo, acetoxy,carb(C₁₋₆)alkoxy, acetonyl, phenyl, (C₁₋₆)alkyl-C(═O)—O—(C₁₋₆)alkyl,(C₁₋₆)alkyl-O—C(═O)—(C₁₋₆)alkyl, (C₁₋₆)hydroxyalkyl, acetamido,(C₁₋₆)alkylcarboxy, (C₁₋₆)alkanoyl, (C₁₋₆)alkyl-keto-(C₁₋₆)alkyl,(C₁₋₆)alkoxy(C₁₋₆)alkyl, napthyl, cyclopropyl, (C₃₋₇)cycloalkyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, benzyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, (C₄₋₈)heteroaryl, glycinyl, alanyl, serinyl,valinyl, leucinyl, isolecinyl, prolinyl, methionyl, phenylalanyl,tyrosinyl, tryptophanyl, threonyl, cystinyl, asparginyl, lysinyl,histidinyl, arginyl, aspartyl, and glutamyl, and wherein the Y isoptionally substituted at a primary nitrogen atom or at a secondarynitrogen atom with the substituent T selected from the group consistingof hydrogen, methyl, (C₁₋₆)alkyl, cyclopropyl, (C₂₋₆)alkenyl,(C₂₋₆)alkynl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, benzyl,(C₄₋₇)cycloalkenyl, phenyl, (C₄₋₁₀)aryl, napthyl, (C₄₋₁₀)haloaryl,(C₄₋₈)heterocyclic, and (C₄₋₈)heteroaryl; and wherein an atom of the Xand an atom of the Y are in a chemical bond that forms a ring,ortho-fused to a ring of Formula (II).
 27. The compound of Formula (II)according to claim 26, wherein the atom of the X and the atom of the Yare in the chemical bond that forms a (C₄₋₈)heterocyclic ring having oneor two heteroatoms selected from the group consisting of N, O, and S,the (C₄₋₈)heterocyclic ring is ortho-fused to a ring of Formula (II),and wherein the X is optionally substituted at the carbon atom with thesubstituent W selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, acetyl, (C₁₋₆)alkyl, nitro, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, and trifluoromethyl, andwherein the X is optionally substituted at the primary nitrogen atom orat the secondary nitrogen atom with the substituent Q selected from thegroup consisting of hydrogen, and (C₁₋₆)alkyl; and wherein the Y isoptionally substituted at the carbon atom with the substituent Zselected from the group consisting of hydrogen, methyl, hydroxy, fluoro,chloro, acetyl, (C₁₋₆)alkyl, nitro, methoxy, (C₁₋₆)alkoxy, thio,methylthio, thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, and trifluoromethyl, and wherein the T is optionallysubstituted at the primary nitrogen atom or at the secondary nitrogenatom with the substituent Q selected from the group consisting ofhydrogen, and (C₁₋₆)alkyl.
 28. The compound of Formula (II) according toclaim 26, wherein the atom of the X and the atom of the Y are in thechemical bond that forms a (C₄₋₈)heteroaryl ring having one or twoheteroatoms selected from the group consisting of N, O, and S, and the(C₄₋₈)heteroaryl ring is ortho-fused to a ring of Formula (II), andwherein the X is optionally substituted at the carbon atom, thesubstituent W selected from the group consisting of hydrogen, methyl,hydroxy, fluoro, chloro, acetyl, (C₁₋₆)alkyl, nitro, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, and trifluoromethyl, andwherein the X is optionally substituted at the primary nitrogen atom orat the secondary nitrogen atom with the substituent Q selected from thegroup consisting of hydrogen, and (C₁₋₆)alkyl; and wherein the Y isoptionally substituted at the carbon atom, the substituent Z selectedfrom the group consisting of hydrogen, methyl, hydroxy, fluoro, chloro,acetyl, (C₁₋₆)alkyl, nitro, methoxy, (C₁₋₆)alkoxy, thio, methylthio,thiomethyl, (C₁₋₆)alkylthio, thio(C₁₋₆)alkyl, chloromethyl,fluoromethyl, and trifluoromethyl, and wherein the T is optionallysubstituted at the primary nitrogen atom or at the secondary nitrogenatom with the substituent Q selected from the group consisting ofhydrogen, and (C₁₋₆)alkyl.
 29. The compound of Formula (II) according toclaim 26, wherein the atom of the X and the atom of the Y are in thechemical bond that forms the ring ortho-fused to a ring of Formula (II),and said ring ortho-fused to a ring of Formula (II) is selected from thegroup consisting of thiophene, pyrrole, furan, imidazole, pyrazole,pyridine pyrazine, pyrimidine, and pyridazine, the ring optionallysubstituted the substituent W selected from the group consisting ofmethyl, hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy, and the ring optionallysubstituted with the substituent Z selected from the group consisting ofmethyl, hydroxyl, fluoro, chloro, bromo, iodo, nitro, cyano, carboxy,carbethoxy, acetyl, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynl, methoxy,(C₁₋₆)alkoxy, thio, methylthio, thiomethyl, (C₁₋₆)alkylthio,thio(C₁₋₆)alkyl, chloromethyl, fluoromethyl, trifluoromethyl,(C₁₋₆)haloalkyl, and (C₁₋₆)haloalkoxy.